Review and Prospectus 2002 - Wellcome Trust Centre For Cell ...

The School of Biological Sciences 

Review and Prospectus 2002 

Wellcome Trust Centre for 

Cell-Matrix Research

contents 

RESEARCH REVIEW 4-5 

Wellcome Trust Centre 

CELL-MATRIX ADHESION AND 6-11 

INTRACELLULAR SIGNALLING 

MATRIX ASSEMBLY AND 12-19 

SUPRAMOLECULAR STRUCTURES 

GENETIC CONTROL OF TISSUE 20-25 

STRUCTURE AND FUNCTION 

PRINCIPAL INVESTIGATOR PROFILES 26-27 

FACILITIES 28 

TRAINING OPPORTUNITIES 29 

STAFF LIST 30 

PUBLICATIONS 31-35 

The Wellcome Trust Centre for Cell-Matrix 

Research is an interdisciplinary research 

centre embedded within the School of 

Biological Sciences at the University of 

Manchester. The Centre was established in 

1995 under the chairmanship of Professor 

Mike Grant with the long-term aims of 

elucidating the structure and function of 

extracellular matrices and cell-matrix 

adhesions, defining the contribution of cellmatrix 

interactions to human diseases, and 

developing approaches for preventing and 

treating these diseases. 

In late 2000, I assumed the position of 

Director of the Centre as Professor Grant was 

first made Dean of the School of Biological 

Sciences, and then Pro-Vice-Chancellor for 

Research of the University. I would like to 

express my personal gratitude to Mike for the 

momentous contributions that he has made to 

cell-matrix research in general, and to the 

matrix group in Manchester in particular, and 

we all wish him well in his new roles. 

Since the publication of our last Review & 

Prospectus in 2000, we have said farewell to 

Sarah Dallas who has moved to a post in the 

Department of Oral Biology at the University of 

Missouri at Kansas City, and in 2002 we will 

see the departure of John Sheehan to the 

Department of Biochemistry at the University 

of North Carolina, to take up the position of 

John Hooker Professor of Biophysics and 

Biochemistry. In the fifteen years that John 

has worked in Manchester, he has pioneered 

the biochemical and biophysical analyses of 

mucins, as well as being instrumental in 

establishing our outstanding facilities for 

purification and analysis of biomolecules. We 

will greatly miss his enthusiasm and creativity. 

By contrast, we welcome Dr. Keith Brennan to 

a Wellcome Trust Research Career 

Development Fellowship in the Centre. Keith 

joins us from the Strang Cancer Research 

Laboratory at Rockefeller University, New York 

to work on Notch and Wnt signalling in 

mammary gland development. We also 

welcome Dr. Clair Baldock to a prestigious 

Royal Society Olga Kennard Research 

page 2

for Cell-Matrix Research 

Fellowship. Clair joined the Centre a few 

years ago as a postdoctoral researcher and 

will now initiate an independent career in the 

area of elastic microfibril structure. Further 

information on Keith and Clair’s work can be 

obtained from two Research in Focus articles 

within this brochure. 

The first twelve months of my period as 

Director have turned out to be most eventful. 

As detailed in this brochure, group leaders 

within the Centre have continued to make 

important discoveries. These advances 

include contributions to the structural biology 

of extracellular matrix molecules and adhesion 

receptors, the biosynthesis and assembly of 

extracellular matrix polymers, and the 

elucidation of genotype-phenotype links in 

diseases caused by mutations in extracellular 

matrix genes. Advances have been made 

across the entire research portfolio of the 

Centre, and have been published in the 

highest quality journals. In addition to 

research output, which is of course of 

paramount importance, I want to highlight 

three other more general achievements that 

augur well for the future prospects of the 

Centre. 

Firstly, in the summer of 2000, we learned that 

an application for re-validation of our status as 

a Wellcome Centre and for renewal of our 

core grant had been successful. The core 

grant will run for five years and provide funds 

for a number of new support posts as well as 

a substantial tranche of new equipment. This 

improvement to our infrastructure will greatly 

facilitate our research and we are extremely 

grateful for the Trust’s support. 

Secondly, in 2000, the School of Biological 

Sciences secured a £15M grant from the 

Wellcome Trust and the UK Government for 

the construction of an Integrative Centre for 

Molecular Cell Biology. This Joint 

Infrastructure Fund award has since been 

combined with further funds from the 

University to create a large complex for 

biological and medical research. The new 

building will be constructed at a focal point in 

the University’s biomedical corridor, adjacent 

to physical sciences research, organismal 

biology research, the pioneering Manchester 

Bioscience Incubator, the Wellcome Trust 

Clinical Research Facility, and the vast Central 

Manchester NHS Trust. The 20,000 m 2 

development will be completed by the end of 

2003 and will provide a new home for our 

Centre in outstanding accommodation. 

Thirdly, in the last few weeks, we have learned 

that the School of Biological Sciences has 

received two top grades of 5* in the Higher 

Education Funding Council for England’s 2001 

Research Assessment Exercise. These 

ratings make the School the largest, top-rated 

Biology department in the UK and will 

determine the level of Government research 

funding to the School for the next five years. 

We anticipate this success enabling the 

School to continue to improve its laboratory 

provision and to recruit outstanding scientists. 

So, as we enter the second five year period of 

the Centre’s existence, the future could hardly 

be brighter. Advances in basic and clinical 

science make this an exciting era for 

biomedical research and we have the real 

prospect of translating our work into practical 

benefits. I hope you enjoy reading about our 

recent progress and future plans in this 

Review & Prospectus. 

Martin Humphries 

page 3

Research Review 

The scientific plans of the Centre can be viewed as three 

integrated and collaborative programmes focused at three 

levels of biological organisation - the cell, the tissue and the 

organism. In brief, the cellular programme focuses on cellextracellular 

matrix adhesion and intracellular signalling, 

and aims to define the molecular principles underlying the 

responses of cells to their environment; the tissue programme 

studies matrix assembly and supramolecular structures 

with the aim of engineering tissues; and the organism 

programme aims to elucidate mechanisms of genetic control 

Cell-matrix adhesion and intracellular 

of tissue structure and function and uses this information for signalling. The overarching aim of this 

improved diagnosis and treatment. Whilst each programme is programme is to decipher the molecular 

mechanisms whereby adhesive cues on the 

highly focused on extracellular matrices and cell-matrix 

outside of cells are transduced into 

interactions, the long-term promise of the work overlaps with functional responses on the inside (and vice 

some of the most important areas of biomedical research - versa) and to develop approaches for 

signalling, tissue engineering and medical genetics. 

intervening experimentally and 

therapeutically in these processes. The 

programme combines strengths in structural 

biology, biochemistry, molecular biology, bioinformatics and cell biology, with an ability to dissect signalling 

pathways in vivo in genetically tractable model organisms such as the mouse, Caenorhabditis, and 

Drosophila. In the long-term, an understanding of the molecular mechanisms and biological functions of 

adhesion will provide insights into tissue formation and homeostasis, and will suggest approaches to 

control many of the most common human diseases, including inflammatory, neoplastic, traumatic and 

infectious conditions. 

Matrix assembly and supramolecular structure. An organised extracellular matrix is essential for the 

function of all tissue and organs. This organisation depends on the cellular control of the co-ordinated 

expression of a range of matrix macromolecules and extracellular assembly into higher order structures. 

In this programme, multidisciplinary approaches are being employed to understand the pathway of 

assembly of extracellular matrices from their initial biosynthesis to the final mature form. The functions of 

different classes of matrix components are being dissected. New approaches to derive structural models 

for higher order macromolecular assemblies, using electron microscopy/tomography, are being combined 

with biophysical analyses, protein crystallography and mass spectrometry to define complex structures. 

These strategies will provide a better understanding of extracellular matrix assembly and the molecular 

basis of the changes associated with skeletal and tissue pathologies. 

Genetic control of tissue structure and function. In this programme, a variety of approaches including 

model vertebrate and invertebrate systems, cell culture systems and human genome scans are being 

employed to gain new insights into the role of the extracellular matrix in determining tissue structure and 

function. Developmental systems under investigation include skeletogenesis, myogenesis, 

vasculogenesis and tissue morphogenesis. Particular progress is being made, firstly, through elucidating 

the molecular pathogenesis of major diseases affecting these organ systems, including osteoarthritis, 

chondrodysplasias and cardiovascular disease, and secondly, by developing new models of diseases 

involving extracellular matrices, such as muscular dystrophies. Understanding the links between the 

individual molecules comprising the extracellular matrix with tissue structure and pathology, is the central 

goal of this research and offers the prospect of developing novel approaches for treatment or prevention 

of several of the most common chronic diseases of mankind. 

page 4

Key Research Discoveries 

Key Research Discoveries 

C.Baldock, K.E.Kadler, C.A.Shuttleworth and 

C.M.Kielty 

The supramolecular organization of fibrillin-rich 

microfibrils. 

J. Cell Biol. 2001; 152:1045-1056. 

M.Baron 

Drosophila dumpy is a gigantic extracellular 

protein required to maintain tension at epidermalcuticle 

attachment sites. 

Curr. Biol. 2000; 10:559-567. 

J.Bella 

Staggered molecular packing in crystals of a 

collagen-like peptide with a single charged pair. 

J. Mol. Biol. 2001; 301:1191-1205. 

P.N.Bishop M.D. Briggs and J.K.Sheehan 

Identification in vitreous and molecular cloning of 

opticin, a novel member of the family of leucinerich 

repeat proteins of the extracellular matrix. 

J. Biol. Chem. 2000; 275:2123-2129. 

R.P.Boot-Handford 

Characterization of Hydra type IV collagen. Type 

IV collagen is essential for head regeneration 

and its expression is up-regulated upon exposure 

to glucose. 

J. Biol. Chem. 2000; 275:39589-39599. 

M.D.Briggs and M.E.Grant 

Mutations in the region encoding the von 

Willebrand factor A domain of matrilin-3 are 

associated with multiple epiphyseal dysplasia. 

Nature Genet. 2001; 28:393-396. 

N.J.Bulleid 

Hsp47: a molecular chaperone that interacts with 

and stabilizes correctly-folded procollagen. 

EMBO J. 2000; 19:2204-2211. 

A.E.Canfield 

1 α,25-dihydroxyvitamin D(3) inhibits 

angiogenesis in vitro and in vivo. 

Circ. Res. 2000; 87:214-220. 

A.P.Gilmore and C.H.Streuli 

Integrin-mediated survival signals regulate the 

apoptotic function of Bax through its 

conformation and subcellular localization. 


M.J.Humphries 

Generation of a minimal α5β1 integrin-Fc 

fragment. 

J. Biol. Chem. 2001; 276:35854-35866. 

K.E.Kadler and C.Baldock 

Corneal collagen fibril structure in three 

dimensions: Structural insights into fibril 

assembly, mechanical properties and tissue 

organization. 

Proc. Natl. Acad. Sci. U. S. A. 2001; 98:7307- 

7312. 

R.A.Kammerer 

The laminin-binding domain of agrin is 

structurally related to N-TIMP-1. 

Nature Struct. Biol. 2001; 8:705-709. 

U.Mayer 

Impaired axonal regeneration in α7 integrindeficient 

mice. 

J. Neurosci. 2000; 20:1822-1830. 

J.K.Sheehan 

Identification of a non-mucin glycoprotein (gp- 

340) from a purified respiratory mucin 

preparation: evidence for an association involving 

the MUC5B mucin. 

Glycobiology. 2001; 11:969-977. 

C.A.Shuttleworth, C.M.Kielty and C.Baldock 

The role of the C1 and C2 A-domains in type VI 

collagen assembly. 

J. Biol. Chem. 2001; 276:7422-7430. 

C.H.Streuli 

Desmosomal adhesion regulates epithelial 

morphogenesis and cell positioning. Nature Cell 

Biol. 2001; 3:823-830. 

D.S.Tuckwell and M.J.Humphries 

Monoclonal antibodies identify residues 199-216 

of the integrin α2 vWFA domain as a functionally 

important region within α2β1. 

Biochem. J. 2000; 350:85-493. 

G.A.Wallis and M.E.Grant 

Coordinated expression of matrix Gla protein is 

required during endochondral ossification for 

chondrocyte survival. 


T.E.Hardingham 

The analysis of intermolecular interactions in 

concentrated hyaluronan solutions suggest no 

evidence for chain-chain association. 

Biochem. J. 2000; 350:329-335. 

page 5

Cell-matrix adhesion and 

intracellular signalling 

MARTIN J. HUMPHRIES 

BSc PhD 

Wellcome Trust Principal 

Research Fellow and Professor of 

Biochemistry 

Key Publications: 

Mould,A.P., J.A.Askari and M.J.Humphries. 

2000. Molecular basis of ligand recognition by 

integrin α5β1. I. Specificity of ligand binding 

is determined by amino acid sequences in the 

second and third NH 2 -terminal repeats of the 

α subunit. J. Biol. Chem. 275:20324- 

20336. 

Humphries,J.D., J.A.Askari, X.P.Zhang, 

Y.Takada, M.J.Humphries and A.P.Mould. 

2000. Molecular basis of ligand recognition by 

integrin α5β1. II. Specificity of Arg-Gly-Asp 

binding is determined by Trp157 of the α 

subunit. J. Biol. Chem. 275:20337-20345. 

Coe,A.P., J.A.Askari,A.D.Kline, M.K.Robinson, 

H.Kirby, P.E.Stephens and M.J.Humphries. 

2001. Generation of a minimal α5β1 

integrin-Fc fragment. J. Biol. Chem. 

276:35854-35866. 

Molecular basis of integrin-dependent 

cell adhesion 

The interactions of integrin adhesion 

receptors with their extracellular matrix 

ligands are important for many aspects of cell 

function. Most notably, they organise 

signalling complexes to modulate 

differentiation and cell fate, provide physical 

support for cells in order to maintain 

cohesion, and permit the generation of 

traction forces to enable movement. Animal 

model studies have also shown integrins to 

contribute to the progression of many 

common diseases, and have implicated them 

as potential therapeutic targets in 

cardiovascular and inflammatory diseases. 

This laboratory is employing a combination of 

biochemical, molecular biological, biophysical 

and cell biological techniques to understand 

the molecular basis of integrin function. The 

aims of this research are to elucidate the 

mode of interaction between integrins and 

their ligands and effectors, usually at the 

atomic level. The work involves narrowing 

down sites within integrin ligands that serve 

as receptor contact sites, determining the 

ligand-binding pocket within integrins, and 

complementing this information through 

tertiary structure determination by X-ray 

crystallography. Such studies have relevance 

for the design of clinically useful anti-adhesive 

agents, and close links with industry are 

facilitating such developments. Recently, the 

regulation of integrin activity has become a 

major emphasis of the research. Integrin 

activation is accomplished through alterations 

in receptor shape that are being probed using 

divalent cations, conformation-dependent 

monoclonal antibodies, and site-directed 

mutagenesis. These studies are now 

translating into investigations of the molecular 

basis of transmembrane signal transduction 

by integrins, in particular integrin-mediated 

organisation of the cytoskeleton through 

agonist-dependent association of cytoskeletal 

proteins. It is hoped that this work will 

provide insights into the mechanisms by 

which cells sense and respond to their 

extracellular environment. 

Co-workers: 

Janet Askari BSc 

Wellcome Trust Technician 

Stephanie Barton BSc 


Mark Bass BSc MRes PhD 

Wellcome Trust Research Associate 

Tanja Benkert Dipl Biol 

Aventis Research Assistant 

Patrick Buckley BSc PhD 


Sue Craig BSc 


Jon Humphries BSc MPhil 

Wellcome Trust Research Assistant 

Lynn McKeown BSc 

BBSRC Student 

Anthea Messent BSc PhD 


Zohreh Mostafavi-Pour BSc MSc PhD 


Paul Mould BSc PhD 


Eileen Pinnington 


Adam Shaw BSc 

BBSRC Student 

Stephen St. George Smith BSc PhD 

BBSRC Research Associate 

Emlyn Symonds BSc 

MRC Student 

Mark Travis BSc 

BBSRC Student 

Dimitra Valdramidou BSc MRes 

Aventis Research Assistant 

Mark Watson BSc 

BBSRC Student 

page 6

JORDI BELLA BSc PhD 

Lecturer in Biochemistry 

Structural biology of cell-matrix proteins 

Key publications: 

Kramer,R.Z., M.G.Venugopal, J.Bella, P.Mayville, 

B.Brodsky, and H.M.Berman. 2000. Staggered 

molecular packing in crystals of a collagen-like 

peptide with a single charged pair. J. Mol. Biol. 

301:1191-1205. 

Kramer,R.Z., J.Bella, P.Mayville, B.Brodsky, and 

H.M.Berman. 1999. Sequence dependent 

conformational variations of collagen triplehelical 

structure. Nat. Struct. Biol. 6:454- 

457. 

Xiao,C., C.M.Bator,V.D.Bowman, E.Rieder,Y.He, 

B.Hebert, J.Bella,T.S.Baker, E.Wimmer, R.J.Kuhn, 

and M.G.Rossmann. 2001. Interaction of 

coxsackievirus A21 with its cellular receptor, 

ICAM-1. J.Virol. 75:2444-2451. 

Co-workers: 

Paul McEwan BSc 

BBSRC Student 

Our current knowledge of the three-dimensional organisation of 

the extracellular matrix and its interaction with cell-surface 

molecules is still rudimentary. My group is particularly interested 

in the structure and function of collagens, as quintessential 

components of all types of extracellular matrices. Collagens are a 

family of proteins characterised by containing at least one domain 

region with a very specific conformation, the collagen triple helix, 

and also by forming higher order macromolecular assemblies. 

Our understanding of the molecular subtleties of the collagen 

triple helix has greatly improved thanks to the crystallographic 

determination of model systems at high resolution. However, key 

questions still remain. For example, we aim to elucidate the basis for molecular recognition of 

collagen, as the triple helix is used as a binding motif for several extracellular macromolecules. 

We will combine crystallographic studies on small versions of collagen with electron microscopy 

reconstructions of entire collagen assemblies. We are also studying the co-assembly of 

collagens with proteins that may control the dimensions or organisation of fibre structures. Major 

efforts are directed at complexes found in cornea and vitreous, and at the molecules mediating 

mineralisation. Finally, we are studying fragments of fibronectin, a molecule that is responsible 

for cell adhesion and migration through its interaction with cell-surface integrins. Solving the 

structures of extracellular matrix proteins, and determining the molecular basis of their 

interaction with other molecules, will contribute to an understanding of their function both in 

normal and diseased states. 

ANDREW P GILMORE BA 

PhD 

Wellcome Trust Research 

Career Development Fellow 


Streuli,C.H. and A.P.Gilmore. 1999.The role of 

adhesion in regulating mammary epithelial cell 

survival. J. Mammary Gland Biol. 

Neoplasia. 4:183-191. 

Gilmore,A.P.,A.D.Metcalfe, L.H.Romer and 

C.H.Streuli. 2000. Integrin mediated survival 

signals regulate the apoptotic function of Bax 

through conformation and subcellular 

localization. J. Cell Biol. 149:431-445. 

Hadjiloucas,I.,A.P.Gilmore, N.J.Bundred and 

C.H.Streuli. 2001.Assessment of apoptosis in 

human breast tissue using an antibody against the 

active form of caspase 3: relation to 

histopathological characteristics. Brit. J. 

Cancer. 85:1522-1526. 

Co-workers: 

Anthony Valentijn BSc 

Wellcome Trust Research Assistant. 

Nadia Zouq BSc MRes 

MRC Student 

Regulation of cell survival 

by cell-extracellular 

matrix adhesion 

Our primary interest lies in extracellular matrix-dependent suppression of apoptosis, which we are 

using as a model for understanding how adhesion-mediated signalling events at the membrane 

are transmitted to other cellular compartments. Apoptosis, or programmed cell death, is a default 

pathway by which damaged or displaced cells are destroyed whilst avoiding a damaging 

inflammatory response, and constant survival signals from the environment are required for cells 

to suppress its activation. The plethora of survival signals are integrated through the Bcl-2 family 

of apoptosis regulators, proteins which function as a gateway between a cells decision to live or 

die. We have found that cell-matrix adhesion activates signalling pathways that suppress the 

activity of Bax, an apoptosis-promoting member of the Bcl-2 family. Bax exerts its death-promoting 

activity at the outer membrane of mitochondria, but in adherent cells adhesion-initiated signals 

keep Bax in the cytosol. Upon inhibition of adhesion, Bax rapidly translocates to mitochondria 

where it initiates apoptosis. 

Our research is currently pursuing two complementary aims. First, we aim to identify the integrinproximal 

signalling pathways that suppress Bax activity in adherent cells, focussing on tyrosine 

kinases activated upon cell adhesion. In addition, the use of GFP-fusions is allowing us to follow 

the movement of Bax in the cytosol of live cells following manipulation of signalling pathways. 

Second, we are examining post-translational modifications of Bax that promote its association with 

mitochondria in cells where the normal adhesion-mediated signalling has been blocked. A variety 

of approaches are being employed, including high-resolution gel filtration and mass spectrometry. 

page 7

CHARLES H STREULI MA PhD 

Wellcome Trust Senior Research 

Fellow 

Adhesion-dependent signalling pathways that 

regulate differentiation and apoptosis 


Gilmore,A.P.,A.D.Metcalfe, L.H.Romer and 

C.H.Streuli. 2000. Integrin mediated survival 

signals regulate the apoptotic function of Bax 

through conformation and subcellular 


Klinowska,T.C., C.M.Alexander, E.Georges- 

Labouesse, R.Van der Neut, J.A.Kreidberg, 

C.J.Jones,A.Sonnenberg and C.H.Streuli. 

2001. Epithelial development and 

differentiation in the mammary gland is not 

dependent on α3 or α6 integrin subunits. 

Dev. Biol. 233:449-467. 

Runswick,S.K., M.J.O’Hare, L.Jones, 

C.H.Streuli and D.R.Garrod. 2001. 


morphogenesis and cell positioning. Nature 

Cell Biol. 3:823-830. 

Co-workers: 

Nasreen Akhtar BSc PhD 


Associate 

Kirsty Green BSc 

BBSRC Student 

Emma Lowe BSc 

HEFCE-funded Technician 

Emma Marshman BSc PhD 

BBSRC Resarch Associate 

QingQiu Pu BS MB PhD 

AstraZeneca Research Associate 

Pengbo Wang BSc MSc PhD 

MRC Research Associate 

Harriet Watkin BSc 

Wellcome Trust Prize Student 

The goal of our research is to understand 

how the extracellular matrix regulates 

epithelial cell phenotype. Matrix proteins 

interact with cell surface integrin receptors to 

modulate cellular architecture and intracellular 

signal transduction pathways. Adhesion to 

basement membranes has a central role in 

controlling the differentiation, apoptosis, and 

morphogenesis of mouse mammary gland 

and human breast epithelium. We are 

dissecting the signalling pathways that control 

these aspects of epithelial cell behaviour. 

Mammary epithelial cells retain their ability to 

express differentiation products in culture, but 

this is dependent on signal transduction 

pathways that are triggered by lactogenic 

hormones and extracellular matrix proteins. 

We have discovered that the signalling 

pathways triggered by integrins and prolactin 

and insulin receptors converge at the level of 

protein tyrosine kinases and protein tyrosine 

phosphatases. A current aim is to identify 

integrin-regulated signalling proteins that 

modulate the pathways driven by lactogenic 

hormones, thereby controlling mammary 

differentiation. 

Breast epithelia undergo rapid and dramatic 

apoptosis after periods of lactation. We are 

dissecting the mechanisms that regulate 

apoptosis, with a particular focus on integrin 

and growth factor control of Bcl-2 family 

proteins. These proteins are associated with 

mitochondrial membranes, and we are 

investigating how two family members, Bax 

and BAD, control apoptotic decisions. We 

have recently found that BAD phosphorylation 

is regulated at the lactation-involution switch 

in vivo, that IGF-BP5 blocks IGF signalling 

and is a potent inducer of apoptosis, and that 

an essential adaptor protein linking IGF 

receptor activation with intracellular signals, 

insulin receptor substrate-1, disappears at 

this time through the activation of a specific 

caspase. Moreover, the ligand-binding 

conformation of the β1 integrin is altered to a 

non-binding state at the onset of apoptosis in 

vivo. 

The morphogenesis of mammary gland 

involves the formation of collecting ducts and 

lactational alveoli, both of which are bilayered 

epithelia. We are investigating how adhesion 

regulates these processes, and have shown 

that mammary gland development in vivo is 

dependent on β1 integrin function. In 

addition, we have identified new functions for 

desmosomal cell-cell adhesion molecules in 

the morphoregulation of alveolus formation. 

We have also discovered that these 

molecules provide essential cues to regulate 

spatial positioning of the epithelial cell types 

in these bilayered structures. 

page 8

DANNY S.TUCKWELL BSc PhD 

BBSRC Advanced Research Fellow 

Modular proteins and the extracellular matrix 


Knight,C.G., L.F.Morton,A.R.Peachey, 

D.S.Tuckwell, R.W.Farndale and M.J.Barnes. 

2000.The collagen-binding A-domains of 

integrins α1β1 and α2β1 recognize the same 

specific amino acid sequence, GFOGER, in 

native (triple-helical) collagens. J. Biol. 

Chem. 275:35-40. 

Tuckwell, D. 2002. Identification and analysis 

of collagen α1(XXI), a novel member of the 

FACIT collagen family. Matrix Biology. 21: 

63-66. 

Aquilina,A., M.Korda, J.M.Bergelson, 

M.J.Humphries, R.Farndale and D.S.Tuckwell. 

2002.A novel gain of function mutation in the 

integrin α2 VWFA domain. European 

Journal of Biochem. in press 

Co-workers: 

Claire Johnston BSc MSc 

BBSRC Student 

The protein components of extracellular 

matrices are typically modular in structure, and 

investigations of the properties of these 

modules can provide insights into the functions 

of the proteins that contain them. Much of the 

research in this laboratory focuses on the 200 

amino acid von Willebrand factor A domain 

module, or VWFA domain, which is found in a 

range of proteins, including integrins, collagens 

and blood coagulation proteins. The 

overarching aim of this work is to determine 

relationships between structure, function and 

evolution within the domain family. Three major 

approaches are being taken: 

1. Integrins: Many cell types interact with 

collagens to regulate normal tissue 

organisation and to mediate more dynamic 

processes such as blood coagulation and 

tumour metastasis. These interactions are 

mediated by members of the integrin family of 

receptors. We have showed that it is the 

VWFA domain within the integrin that binds to 

collagens, and recent work has characterised 

the molecular basis of this interaction. 

Currently, we are studying the interaction of the 

α2 VWFA domain with non-collagenous ligands 

and naturally occurring inhibitors, with a view to 

developing novel therapeutic agents. 

2. Caenorhabditis elegans: Bioinformatics 

analysis shows that there are 33 VWFA 

domain-containing proteins in the nematode C. 

elegans. We have used an integrative 

approach to study the novel VWFA domain 

protein DDA-1 and shown that it is required for 

the correct formation of the cuticle. We now 

intend to use genetic and biochemical 

approaches to identify the proteins interacting 

with DDA-1, thereby linking matrix genes with 

specific functions within the organism. 

3. Bioinformatics and phylogeny: Much of our 

work employs bioinformatics approaches to 

complement and extend lab-based studies, 

including structure prediction, homology 

modelling and phylogenetic analysis. We 

recently identified a novel human collagen 

[collagen α1(XXI)] using bioinformatics tools, 

and showed that it diverged early in vertebrate 

collagen evolution. Over the next few years, 

bioinformatics methodologies will be used to 

elucidate the functional evolution of other 

modules, including vertebrate collagen C- 

propeptides and C. elegans cuticle ZP 

domains. 

In conclusion, our studies have resulted in the 

detailed characterisation of known interactions 

and the discovery of novel functions. VWFA 

domains will continue to be a rewarding field for 

research; moreover, the strategies outlined 

above will also facilitate the exploration of other 

domain families. 

page 9

MARTIN BARON BSc PhD 

Lecturer in Biochemistry 


Fostier,M., D.Evans, S.Artavanis-Tsakonas and 

M.Baron. 1998. Genetic characterisation of the 

Drosophila melanogaster Suppressor of deltex:A 

regulator of Notch signalling. Genetics 

150:1477-1485. 

Cornell,M., D.Evans, R.D.Mann, M.Fostier, 

M.Flasza, M.Monthatong, S.Artavanis-Tsakonas 

and M.Baron. 1999 The Suppressor of deltex gene 

a regulator of the Notch receptor signalling 

pathway is an E3 class ubiquitin ligase. Genetics 

152:567-576. 

Baron,M.,V.O’Leary, D.A.P.Evans, M.Hicks and 

K.Hudson. 2000. Multiple Roles of the Dcdc42 

GTPase during wing development in Drosophila 

melanogaster. Molecular and General 

Genetics 264:98-104. 

Regulation of Notch receptor signalling 

The control of cell differentiation during development requires communication between cells 

via diffusible growth factors and cell-cell adhesion mediated signalling molecules. The Notch 

receptor is an example of the latter class and regulates the timing and outcome of cell 

differentiation decisions in many different tissues 

during development. The Notch signal must be 

precisely regulated to prevent inappropriate signalling. 

Notch was first identified in Drosophila but has 

subsequently been identified in a range of vertebrate 

species, including four versions of the gene in 

humans. The activity of Notch in vivo is precisely 

controlled spatially and temporally. Using the 

Drosophila model system, our group has identified a 

negative regulator of Notch called Suppressor of 

Deltex [Su(dx)], which belongs to an E3 class of 

ubiquitin ligase molecules that regulate endocytosis 

and proteolytic degradation of target molecules. We 

are currently investigating the mechanism of Notch 

pathway regulation by Su(dx) using a combination of 

genetic, biochemical and cell biological approaches. 

Co-workers: 

Ann-Marie Carbery BSc MSc, BBSRC Student 

Maggy Fostier BSc PhD, Wellcome Trust Research Associate 

Jenny Higgs BSc, BBSRC Student 

Sabine Mazaleyrat BSc, BBSRC Student 

Ngoc-sa Nguyen Huu BSc MSc, MRC Research Assistant 

Marian Wilkin BSc PhD, MRC Research Associate 

KEITH R. BRENNAN BA PhD 



Interactions between signalling pathways 

during mammalian development 


Brennan,K., M.Baylies and A.Martinez Arias. 

1999.A repressive function of Notch required 

prior to Wingless signalling during muscle 

progenitor cell development in Drosophila. 

Current Biology. 9:707-710. 

Brennan,K.,T.Klein, E.Wilder and A.Martinez 

Arias 1999. Wingless modulates the effects of 

dominant negative Notch molecules in the 

developing wing of Drosophila. Implications for 

Notch/Wingless signalling. Developmental 

Biology. 216:210-229. 

Lawrence,N.,T.Langdon, K.Brennan and 

A.Martinez Arias. 2001. Notch signalling targets 

the Wingless response element of a Ubx enhancer 

in Drosophila. Current Biology. 11:375-385. 

The interactions between signalling pathways are crucial for their regulation during embryonic 

development and the maintenance of adult tissues, and disruption of these feedback mechanisms 

can lead to developmental defects and disease. The Notch and Wnt pathways are two of the most 

important developmental signalling pathways and are required for multiple aspects of mammalian 

development. Frequently, signalling through both pathways is required for the formation of an organ 

although they typically have opposing effects on cell fate decisions. Through detailed genetic 

analysis in Drosophila, and recently in mammals, we have highlighted a novel role of Notch in the 

repression of Wnt target genes prior to the receipt of a Wnt signal. This function does not require 

known elements of the Notch signalling pathway, although 

recent experiments suggest that the cytoplasmic protein Deltex 

is required. Current experiments seek to elucidate the 

molecular mechanism that underlies this regulatory interaction. 

Continuation of the cell culture experiments, using established 

Wnt and Notch signalling assays and cell lines whose 

differentiation is regulated by both pathways, will identify the 

point(s) of regulation and the proteins involved. In parallel, 

interactions between the pathways will be examined in vivo 

within the developing mammary gland where signalling through 

both pathways is required and intersects. Transgenic mice and 

conditional knockouts will be used to manipulate Wnt and 

Notch signalling. Careful examination of the resulting 

mammary gland phenotypes will reveal whether signalling 

through one pathway can regulate the other. 

page 10

Research in Focus 

Notch signal regulation and cell proliferation 

In defining a body plan and regulating cellular 

responses to positional cues, organisms do 

not use a different signal for each individual 

cell-fate decision, but repeatedly use a 

relatively small set of cellular signalling 

pathways. The Notch receptor and its 

membrane bound ligands, Delta and Serrate, 

were first identified in Drosophila and are vital 

components of one such developmentally 

important signalling pathway. Four 

homologues of Notch are found in humans 

and aberrant Notch signalling levels have 

been linked to diverse human diseases 

including neural degenerative diseases, 

developmental disorders and cancer, making 

this pathway an important target for 

therapeutic interventions. 

Notch is best known for its role in lateral inhibition 

signalling where a small number of cells are selected 

to adopt a particular fate from a larger group of 

precursor cells. This lateral inhibition signalling 

depends on a ligand-dependent cleavage of Notch, 

which allows its intracellular domain to activate a 

transcriptional regulator, Suppressor of Hairless 

[Su(H)], in the nucleus. However, recent work has 

indicated that there is a second intracellular signalling 

pathway, which requires the Notch interacting protein 

Deltex, acting downstream of the receptor. 

The regulation of Notch signalling occurs at 

many levels, including cross-talk with other 

signalling pathways, covalent modifications 

including glycosylation, phosphorylation and 

ubiquitination, and intracellular trafficking of 

the receptor and ligands. Our aim is to 

unravel these intersecting mechanisms of 

regulation. 

One mechanism of regulation under 

investigation is the role played by ubiquitin 

ligase proteins which covalently modify target 

proteins and regulate their intracellular 

trafficking and turnover. Several Notch 

regulatory proteins have been identified 

which have either been shown to act as 

ubiquitin ligases or contain ubiquitin ligaserelated 

domains. In our laboratory, a 

negative regulator of Notch signalling, 

Suppressor of deltex [Su(dx)], has been 

identified that belongs to the E3 class of 

ubiquitin ligases. Like Deltex, this protein 

associates with the Notch intracellular 

domain. However, it is currently unclear how 

Su(dx) regulates Notch signalling and 

whether regulation occurs through Su(dx)- 

dependent ubiquitination of Notch. In 

addition, genetic experiments indicate that 

Su(dx) and Deltex may interact to regulate 

certain aspects of Notch signalling. 

Understanding the mechanism of action of 

these proteins and their relationship to the 

two intracellular signalling pathways 

downstream of Notch are important goals of 

our research. 

A second important avenue of research is the 

investigation of the cross-talk between the 

Notch signalling pathways and another 

developmentally important signal, the 

Wingless (Wnt-1 in vertebrates) pathway. 

Our genetic experiments indicate that Notch 

signalling through the Deltex-dependent 

pathway suppresses Wingless target genes 

prior to the receipt of a Wingless signal in 

Drosophila. In addition, our results suggest 

that an important step in Wingless signalling 

is to attenuate Notch signalling via Deltex. 

Finally, preliminary experiments indicate that 

regulatory interactions occur between the 

Notch and Wnt pathways in mammals. 

Understanding the molecular basis for this 

regulatory interaction will give insights into 

how these two pathways collaborate to 

control developmental patterning. 

In summary, our research has highlighted two 

distinct mechanisms for regulating Notch 

signalling. By using a wide variety of genetic, 

cell biological and biochemical methods we 

are currently unravelling the molecular 

mechanisms behind these regulatory 

interactions. Finally we are translating our 

results from the model organism, Drosophila, 

to human disease. 

Martin Baron and Keith Brennan 

page 11

Matrix Assembly and 

Supramolecular Structures 

KARL E. KADLER BSc PhD 

Professor of Biochemistry 


Graham,H.K., D.F.Holmes, R.B.Watson and 

K.E.Kadler. 2000. Identification of collagen 

fibril fusion during vertebrate tendon 

morphogenesis.The process relies on unipolar 

fibrils and is regulated by collagenproteoglycan 

interaction. J. Mol. Biol. 

295:891-902. 

Garrigue-Antar,L., C.Barker and K.E.Kadler. 

2001. Identification of amino acid residues in 

bone morphogenetic protein-1 important for 

procollagen C-proteinase activity. J. Biol. 

Chem. 276:26237-26242. 

Holmes,D.F., C.J.Gilpin, C.Baldock, U.Ziese, 

A.J.Koster and K.E.Kadler. 2001. Corneal 

collagen fibril structure in three dimensions: 

Structural insights into fibril assembly, 

mechanical properties and tissue organization. 

Proc. Natl.Acad. Sci. U. S.A 98:7307- 

7312. 

The molecular and cellular basis of 

collagen fibrillogenesis 

My laboratory aims to determine the 

molecular and cellular basis of collagen 

fibrillogenesis during tissue assembly. Tissue 

assembly in animals requires the formation of 

an extracellular matrix containing millimetrelong 

collagen fibrils arranged in elaborate 

three-dimensional architectures such as 

parallel bundles (in tendons and ligaments), 

basket weaves (in skin and bone) and 

orthogonal lattices (in cornea). We are taking 

a multidisciplinary approach to studying 

collagen fibrillogenesis, by using a 

combination of electron microscopy (to 

determine the three-dimensional structure of 

individual collagen fibrils), site-directed 

mutagenesis and recombinant protein 

expression (to understand how collagen is 

synthesised), and immunolocalisation 

techniques (to study protein trafficking leading 

to collagen fibrillogenesis). Our work has 

direct relevance to protein trafficking of large 

and multimeric proteins, and in understanding 

the aetiology of some of the most debilitating 

diseases of man and animals, such as 

fibrosis and arthritis, that are characterised by 

ectopic and unusual synthesis of collagen 

fibrils. 

Analysis of the synthesis of large multimeric 

polymers, that are orders of magnitude longer 

than the cells that produce them, is difficult 

because they are refractory to conventional 

approaches. We have used transmission 

electron microscopy, which has led to the 

discovery that the fibrils are synthesised as 

two types of ‘collagen early fibrils’ that have 

the ability to fuse (end-to-end and end-toside) 

to generate long fibrils that, in turn, 

anastomose and form branched networks in 

developing tissues. Recently, we have used 

automated electron tomography and 3D 

reconstruction to obtain the first threedimensional 

structure of individual collagen 

fibrils. A new focus of interest is to understand 

how the trafficking of key proteins involved in 

fibrillogenesis, including procollagen and 

bone morphogenetic protein-1, influences 

collagen fibril assembly, as well as the 

structure and function of the extracellular 

matrix. 

Co-workers: 

Adetola Adesida BSc PhD 

Wellcome Trust/Catalyst Biomedica Ltd 

Research Associate 

Elizabeth Canty BSc PhD 


Laure Garrigue-Antar BSc PhD 


Hanane Gouizi BSc 

Algerian Government Student 

Nicola Hartigan BSc 


David Holmes BSc PhD 


Matthew Leighton BSc PhD 


Roger Meadows BSc MSc 

Wellcome Trust technician 

Vasiliki Petropolou BSc MSc 

University of Manchester Student 

Susan Richardson BSc PhD 

Wellcome Trust/Catalyst BioMedica Ltd 

Research Associate 

Tobias Starborg BSc PhD 

BBSRC/MRC/EPSRC Research Associate 

page 12

NEIL J. BULLEID BSc PhD 



Tasab,M., M.R.Batten and N.J.Bulleid. 2000. 

Hsp47: a molecular chaperone that interacts with 

and stabilizes correctly-folded procollagen. 

EMBO J. 19:2204-2211. 

Wilson,C.M., M.R.Farmery and N.J.Bulleid. 

2000. Pivotal role of calnexin and mannose 

trimming in regulating the endoplasmic 

reticulum-associated degradation of major 

histocompatibility complex class I heavy chain. J. 

Biol. Chem. 275:21224-21232. 

Bottomley,M.J., M.R.Batten, R.A.Lumb and 

N.J.Bulleid. 2001. Quality control in the 

endoplasmic reticulum. PDI mediates the ER 

retention of unassembled procollagen C- 

propeptides. Curr. Biol. 11:1114-1118. 

Co-workers: 

Claire Bithell BSc 

BBSRC Student 

Seema Chakravarthi BSc MSc 


Alexandra Hillebrand BSc 

Marie Curie Visiting Student 

Catherine Hoare BSc MSc 


Lynsey Jenkinson BSc 


Richard Lumb BSc 

BBSRC Student 

Pooli Ragesekariah BSc PhD 


Mohammed Tasab BSc PhD 


Mark Warren BSc PhD 


Rachel Watkins BSc 


The maturation and secretion of procollagen 

A major aim of the research in this laboratory is to elucidate the fundamental molecular 

interactions that control the biosynthesis and folding of extracellular matrix proteins. The lumen 

of the endoplasmic reticulum contains a number of proteins that have been shown previously to 

interact with polypeptide chains during their maturation and transport through the ER to the 

Golgi apparatus. These proteins include enzymes that catalyse folding events and molecular 

chaperones that interact with folding polypeptide chains to prevent premature aggregation or 

non-specific interactions. 

Procollagen is an excellent example of a molecule that interacts with a number of enzymes and 

molecular chaperones during its folding and assembly. If the initial folding of the procollagen is 

prevented, the unfolded chains associate with BiP, which eventually leads to proteasomemediated 

degradation. Once the C-propeptide domains have folded, they either assemble to 

form trimers immediately or associate with protein disulfide isomerase until interacting chains are 

synthesised. Folding of the triple helical domain follows this trimerisation event. Trimeric, nontriple 

helical molecules interact with prolyl-4-hydroxylase (P4H), an interaction that depends on 

the folding status of the protein rather than hydroxylation state. Once the triple helix has formed, 

the protein is transported to the Golgi apparatus where it forms higher order aggregates 

resulting in characteristic distensions of the Golgi cisternae. 

This is not the complete picture, however, as procollagen chains have also been shown to 

interact with a heat shock protein (Hsp47). What role could this protein be playing in 

procollagen biosynthesis? Clearly it is an essential protein as mice deficient in Hsp47 die before 

birth. We are currently investigating a variety of different hypotheses as to the role of this novel 

molecular chaperone, which is one of the few examples of a protein-specific chaperone. 

page 13

TIMOTHY E HARDINGHAM 

PhD DSc 


Key publications; 

Day,J.M.,A.D.Murdoch, and T.E.Hardingham. 

1999.The folded protein modules of the C- 

terminal G3 domain of aggrecan can each 

facilitate the translocation and secretion of the 

extended chondroitin sulfate attachment sequence. 

J. Biol. Chem. 274:38107-38111. 

Gribbon,P., B.C.Heng, and T.E.Hardingham. 

2000.The analysis of intermolecular interactions 

in concentrated hyaluronan solutions suggest no 

evidence for chain-chain association. Biochem. J. 

350 Pt 1:329-335. 

Kolettas,E., H.I.Muir, J.C.Barrett, and 

T.E.Hardingham. 2001. Chondrocyte phenotype 

and cell survival are regulated by culture 

conditions and by specific cytokines through the 

expression of Sox-9 transcription factor. 

Rheumatology. (Oxford) 40:1146-1156. 

Co-workers: 

Chris Brew BSc MBChB MRCS 

ARC Clinical Training Fellow 

Alan Murdoch BSc PhD 

BBSRC/MRC/EPSRC Research 

Fellow 

Janine Prince BSc MSc 

BBSRC/MRC/EPSRC Technician 

Richard Rauchenberg BSc 

BBSRC Student 

Bertrand Raynal BSc MSc 


Simon Tew BSc PhD 


Associate 

Proteoglycan functions in the extracellular matrix 

The formation, turnover and repair of the extracellular matrix are essential determinants of 

the growth and development of tissues into organs. Proteoglycans are key components of 

the extracellular matrix, and aggrecan is the major proteoglycan of cartilaginous tissue. 

Aggrecan, which comprises a multi-domain protein core attached to a large number of 

chondroitin sulphate and keratan sulphate glycosaminoglycan chains, forms supramolecular 

aggregates by binding to hyaluronan. We are using biochemical, molecular biology and 

biophysical techniques to investigate the protein domain and glycosaminoglycan functions of 

aggrecan. A new approach using fluorescence recovery after photobleaching with a confocal 

microscope (confocal-FRAP) has been developed to study molecular interactions of 

proteoglycans, hyaluronan and other glycoconjugates, such as mucins. With this technique, 

which employs high molecular concentrations close to those found physiologically, the 

networks formed by hyaluronan and aggrecan in solution and their permeability to FITClabelled 

probes are being investigated. This approach is also being applied to sections of 

cartilage to follow permeability changes that accompany matrix damage and new matrix 

assembly, and we are studying the changes in gene expression in chondrocytes that 

characterise osteoarthritis. 

We also have an active programme in cartilage tissue engineering. Chondrocytes are 

responsible for the production of cartilage by extracellular matrix, but lose the ability to make 

extracellular matrix in cell culture. We are investigating the expression of SOX genes in 

human chondrocytes during the loss of phenotype in cell culture and the effects of 

transfected SOX genes on the recovery of phenotype. The synthesis and assembly of 

cartilage matrix by chondrocytes in culture will be assessed by composition analysis, 

confocal microscopy, and by confocal-FRAP analysis to monitor matrix network formation. 

This will identify the key interactions in the initial assembly of extracellular matrix by 

chondrocytes and will lead to new strategies for generating cartilage matrix in culture. 

page 14

CAY M. KIELTY BSc PhD 

MRC Senior Research Fellow 

and Professor of Medical 

Biochemistry 

ADRIAN SHUTTLEWORTH 

BSc PhD 

Reader in Medical Biochemistry 

The structure and function of extracellular matrix 

macromolecules in health and disease 


Baldock,C.,A.J.Koster, U.Ziese, M.J.Rock, 

M.J.Sherratt, K.E.Kadler, C.A.Shuttleworth, and 

C.M.Kielty. 2001.The supramolecular 

organization of fibrillin-rich microfibrils. J. Cell 

Biol. 152:1045-1056. 

Ball,S.G., C.Baldock, C.M.Kielty, and 

C.A.Shuttleworth. 2001.The role of the C1 and 

C2 a-domains in type VI collagen assembly. J. 

Biol. Chem. 276:7422-7430. 

Chaudhry,S.S., J.Gazzard, C.Baldock, J.Dixon, 

M.J.Rock, G.C.Skinner, K.P.Steel, C.M.Kielty and 

M.J.Dixon. 2001. Mutation of the gene encoding 

fibrillin-2 results in syndactyly. Hum. Mol. 

Genet. 10:835-843. 

Our aims are to understand how cells regulate the formation of supramolecular assemblies in 

health and disease, and how such assemblies are organised within, and contribute functionally 

to, diverse extracellular matrices. We are correlating genotype with phenotype in heritable 

connective tissue disorders, and investigating the processes underlying vascular matrix 

remodelling. Molecular, cellular and microscopy approaches are being applied to define the 

structure and function of three major supramolecular assemblies of the extracellular matrix. 

1. Fibrillin-rich microfibrils are complex, extensible polymers that play a key role in elastic fibre 

formation. Their essential contribution to connective tissue integrity is highlighted by linkage of 

mutations in fibrillin genes to Marfan syndrome and related diseases associated with 

cardiovascular, skeletal and ocular defects. We are expressing fibrillin in recombinant form to 

investigate its assembly and molecular interactions, and using electron microscopy and atomic 

force microscopy to study microfibril organisation and elasticity. 

2. Type VI collagen microfibrils play a key linking role in the extracellular matrix. Mutations in 

this collagen cause the heritable skeletal muscle disorder, Bethlem myopathy. We are 

investigating the unique hierarchical assembly of type VI collagen microfibrils by analysing the 

specific interactions between its triple helix and von Willebrand factor A-domains, and identifying 

genotype-to-phenotype correlations that affect microfibril assembly. 

3. Type VIII collagen is a structural component of the vasculature that is upregulated following 

vascular injury and during remodelling. Mutations in this collagen cause two forms of corneal 

endothelial dystrophy. Type VIII collagen forms hexagonal lattices that may provide pericellular 

structural support for proliferating vascular smooth muscle cells and endothelial cells. 

Recombinant expression approaches are being used to examine its chain composition, 

supramolecular assembly and biological interactions. 

Co-workers: 

Craig Barley BSc 

ARC Research Assistant 

David Murray MBChB MRCS 

BBSRC/MRC/EPSRC Clinical Training Fellow 

Sarah Bernard BSc 

BBSRC Student 

Steve Ball BSc PhD 


Dan Bax BSc 

BHF Research Associate 

Claire Crouchley BSc 

BBSRC/MRC/EPSRC Student 

Nigel Hodson BSc PhD 

MRC Co-op Research Associate 

Karen Johnston BSc PhD 


David Lee BSc PhD 


Amanda Lomas BSc 

BBSRC/MRC/EPSRC Technician 

Kieran Mellody BSc MSc 


Amanda Morgan BSc 


Suzannah Phillips BSc 

BBSRC Student 

Gareth Pugh BSc PhD 


Matt Rock BSc PhD 


Mike Sherratt BSc PhD 


Simon Stephan BSc PhD 


page 15

PAUL N. BISHOP 

PhD FRCS FRCOphth 

Wellcome Trust Senior Research 

Fellow in Clinical Science 


Reardon,A.J., M.Le Goff, M.D.Briggs, D.McLeod, 

J.K.Sheehan, D.J.Thornton and P.N.Bishop. 2000. 

Identification in vitreous and molecular cloning of 

opticin, a novel member of the family of leucinerich 

repeat proteins of the extracellular matrix. 

J. Biol. Chem. 275:2123-2129. 

Bos,K.J., D.F.Holmes, K.E.Kadler, D.McLeod, 

N.P.Morris and P.N.Bishop. 2001.Axial structure 

of the heterotypic collagen fibrils of vitreous 

humour and cartilage. J. Mol. Biol. 306:1011- 

1022. 

Takanosu,M.,T.C.Boyd, M.Le Goff, S.P.Henry, 

Y.Zhang, P.N.Bishop and R.Mayne. 2001. 

Structure, chromosomal location and tissue-specific 

expression of the mouse opticin gene. 

Invest.Ophthalmol.Vis. Sci. 42:2202-2210. 

Co-workers: 

Jane Bradley BSc 


Magali Le Goff BSc MSc 


V. John Hindson BSc PhD 


Tom Jowitt BSc 

Iris Fund Research Associate 

Lisa Macrory BSc 


Wang Jing MD 

Clinical Research Fellow 

Matrix biology of the eye 

The gel state of the vitreous humour is maintained by a loose network of heterotypic collagen fibrils, containing collagens types II, IX, 

and V/XI. These collagen fibrils are coated with non-collagenous structural macromolecules and we hypothesise that these 

molecules also play a key role in the supramolecular organisation of the vitreous gel. A major focus of our research has been to 

understand how these collagenous and non-collagenous components are organised within the vitreous humour and how this 

organisation is perturbed in ageing and disease. Using electron microscopy combined with computer modelling, we have elucidated 

the axial distribution of collagen molecules within the fibrils. We have also determined how the collagen fibrils are distributed through 

the vitreous gel and the effects of ageing upon these different levels of organisation. We isolated and characterised a pool of 

collagen-associated molecules from the vitreous gel and this work led to the discovery of a novel member of the extracellular matrix 

small leucine-rich repeat protein family that we have named opticin. Opticin expression is virtually restricted to the non-pigmented 

ciliary epithelium of the eye and the onset of expression coincides with ciliary body differentiation, thereby providing a marker of this 

developmental process. Opticin is highly expressed by both the embryonic and adult eye and we are currently elucidating its 

functions using a combination of genetic, biochemical and cell biological strategies. 

page 16

JOHN K SHEEHAN 

BSc MSc PhD 

Reader in Physiological 

Biochemistry 

The structure and function of mucus 

DAVID J.THORNTON BSc PhD 

Senior Experimental 

Officer/Senior Research 

Scientist 


Sheehan,J.K., C.Brazeau, S.Kutay, H.Pigeon, 

S.Kirkham, M.Howard and D.J.Thornton. 2000. 

Physical characterization of the MUC5AC mucin: 

a highly oligomeric glycoprotein whether isolated 

from cell culture or in vivo from respiratory 

mucous secretions. Biochem. J. 347: 37-44. 

Thornton,D.J.,T.Gray, P.Nettesheim, M.Howard, 

J.S.Koo and J.K.Sheehan. 2000. Characterization 

of mucins from cultured normal human 

tracheobronchial epithelial cells. Am. J. Physiol. 

Lung Cell Mol. Physiol. 278:L1118-L1128. 

Thornton,D.J., J.Davies, S.Kirkham,A.Gautrey, 

N.Khan, P.S.Richardson and J.K.Sheehan. 2001. 

Identification of a non-mucin glycoprotein (gp- 

340) from a purified respiratory mucin 

preparation: evidence for an association involving 

the MUC5B mucin. Glycobiology. 11: 969- 

977. 

Our work is centred on understanding the supramolecular assembly 

and function of mucus gels, with a specific interest in the role of 

mucus in the human airways in health and disease. Mucus is central 

to the protection and the maintenance of homeostasis of the lung. 

This highly hydrated gel, in conjunction with ciliated epithelial cells, 

forms the mucociliary escalator, which along with cough, is essential 

for the sterility of the airways. By contrast, overproduction of mucus 

with altered physical properties is an important factor in the morbidity 

and mortality of chronic airways disease such as asthma, cystic 

fibrosis and chronic obstructive pulmonary disease. Our work until 

recently has centred upon the identification, characterisation and 

quantitation of the large oligomeric O-linked glycoproteins (mucins) 

responsible for the properties of the mucus gels found in healthy and 

diseased airways secretions. We have been successful in devising 

probes to identify distinct mucin species secreted from different 

cellular sources in the respiratory tract (i.e. from the surface epithelia 

or underlying submucosal glands). It has become clear that the 

mucins are the organising framework around which many protective 

molecules are condensed. In the coming phase of our work we are 

seeking to understand the higher-order organisation of mucins and 

the role other proteins play in mucus structure. A large part of the 

organisation arises from the initial mechanisms of biosynthesis and 

packaging of the mucins into intracellular storage granules prior to 

their secretion and thus our interests in these processes are 

expanding. We are also pursuing a vigorous proteomics approach 

for the identification and quantitation of other proteins and 

glycoproteins involved in mucus organisation and at the same time 

are devising new methods and strategies for assaying the network 

properties of the mucus gel itself. 

Co-workers: 

Linda Eyers BSc 

BBSRC Student 

Sara Kirkham BSc 


Matthew Wakefield BSc 


page 17

RICHARD A. KAMMERER 

Dipl Biochemistry PhD 




Kammerer,R.A.,V.A.Jaravine, S.Frank, 

T.Schulthess, R.Landwehr,A.Lustig, C.Garcia- 

Echeverria,A.T.Alexandrescu, J.Engel, and 

M.O.Steinmetz. 2001.An intrahelical salt bridge 

within the trigger site stabilizes the GCN4 leucine 

zipper. J. Biol. Chem. 276:13685-13688. 

Stetefeld,J., M.Jenny,T.Schulthess, R.Landwehr, 

J.Engel, and R.A.Kammerer. 2000. Crystal 

structure of a naturally occurring parallel righthanded 

coiled coil tetramer. Nat. Struct. Biol. 

7:772-776. 

Coiled coils in extracellular matrix proteins 

The goal of our research is to improve our understanding of protein-protein interactions in 

extracellular matrices in order to understand the processes determining their formation and 

their disruption in disease. We are using the coiled-coil element as a model system to 

study these interactions. Coiled coils mediate subunit oligomerisation in a large number of 

proteins and are implicated in a wide variety of biological functions. The left-handed coiledcoil 

motif is a type of protein structure consisting of two to five right-handed amphipathic α- 

helices that “coil” around each other in a slight super-twist. The sequences of left-handed 

coiled coils are characterized by a heptad repeat of seven residues denoted a to g with a 

3,4-hydrophobic repeat of mostly apolar amino acids at positions a and d. The helices in 

coiled-coil assemblies can be arranged in a parallel or anti-parallel manner. In addition, 

coiled-coil interactions can result in homo- or heterotypic oligomers. The simplicity of its 

structure makes the coiled-coil structural motif an attractive system for studying both the 

intra- and inter-molecular interactions that govern the folding and stability of multi-subunit 

proteins. Using a combination of molecular biology, biochemistry, biophysical chemistry, 

and structural biology, our efforts are aimed at understanding these interactions. 

Stetefeld,J., M.Jenny,T.Schulthess, R.Landwehr, 

B.Schumacher, S.Frank, M.A.Ruegg, J.Engel, and 

R.A.Kammerer. 2001.The laminin-binding 

domain of agrin is structurally related to N- 

TIMP-1. Nat. Struct. Biol. 8:705-709. 

Co-workers: 

Philip Macdonald BSc 


CLAIR BALDOCK BSc PhD 

Royal Society Olga Kennard 

Research Fellow 

Structural studies on microfibrillar 

components of the 

extracellular matrix 


Baldock,C.,A.J.Koster, U.Ziese, M.J.Rock, 

M.J.Sherratt, K.E.Kadler, C.A.Shuttleworth and 

C.M.Kielty. 2001.The supramolecular 

organization of fibrillin-rich microfibrils. J. Cell 

Biol. 152:1045-1056. 

Ball,S.G., C.Baldock, C.M.Kielty and 

C.A.Shuttleworth. 2001.The role of the C1 and 

C2 A-domains in type VI collagen assembly. J. 

Biol. Chem. 276:7422-7430. 





mechanical properties and tissue organization. 

Proc. Natl.Acad. Sci. U. S.A 98:7307- 

7312. 

The aim of the research in this laboratory is to investigate 

the structure and function of microfibrillar components of 

the extracellular matrix using a combination of electron 

microscopy and X-ray crystallography. Two different 

microfibrillar assemblies are being studied - the fibrillin-rich 

microfibrils and collagen VI microfibrils - each of which 

makes an essential contribution to normal tissue elasticity. 

Resolving extracellular matrix polymers using conventional structural biology techniques has 

previously proved difficult due to their size and complexity, but now these problems can be 

circumvented with the new technique of electron tomography. By marrying cryo-electron 

microscopy data with high-resolution structures of key domains, high resolution information can be 

extrapolated to complex extracellular assemblies. 

The goal of this research is to provide new insights into the molecular architecture of these large 

and ultrastructurally complex microfibrils. This structural information will provide a better 

understanding of their biological properties in health and disease. 

Co-workers: 

Andrew Marson BSc 

BBSRC Student 

page 18


The supramolecular organisation of fibrillin-rich microfibrils 

The elastic fibre system makes a key 

contribution to the structure and function of 

organs that require elasticity, such as large 

arteries, lung and skin. The principal 

components of elastic fibres that endow 

them with their special physical properties 

are polymerised elastin and fibrillin-rich 

microfibrils. The major aims of our research 

are to determine the mechanisms of 

assembly and organisation of fibrillin-rich 

microfibrils, and to elucidate their role in 

elastic fibre formation. 

Fibrillin-rich microfibrils are a unique class of 

multicomponent extracellular matrix microfibril that 

endow connective tissues with long-range elasticity. 

In the untensioned state, they have a repeating 

periodicity of 56nm with a ‘beads-on-a-string’ 

appearance and a diameter of 15nm. Their principal 

structural molecule is fibrillin, a large cysteine-rich 

glycoprotein (~350 kDa) that forms the molecular 

scaffold of this class of microfibril. The importance 

of fibrillin-rich microfibrils is emphasised by the 

linkage of fibrillin mutations to Marfan syndrome and 

related connective tissue disorders that are 

associated with severe cardiovascular, ocular and 

skeletal defects. Until now, however, the 

arrangement of fibrillin molecules within microfibrils 

has remained poorly defined. 

Automated electron tomography was used 

to generate 3D microfibril reconstructions, 

which revealed many new organisational 

details of untensioned microfibrils, including 

heart-shaped beads from which two arms 

emerge, and inter-bead diameter variation. 

The reconstructions revealed that 

microfibrils comprise two in-register 

filaments with a longitudinal symmetry axis, 

with up to eight fibrillin molecules in crosssection. 

Antibody epitope mapping of untensioned 

microfibrils revealed the overlap of epitopes 

at the N- and C-terminus and the 

juxtaposition of two internal epitopes that 

would be 42 nm apart in unfolded 

molecules, which infers intramolecular 

folding. Comparison of colloidal gold and 

antibody binding sites in untensioned 

microfibrils and those extended in vitro 

highlight conformational changes and 

intramolecular folding. 

Together our data indicate that an ~onethird 

stagger is adopted in untensioned 

microfibrils, but a molecular head-to-tail 

arrangement could occur in highly extended 

microfibrils. Our model suggests a 

molecular basis for microfibril extensibility 

which involves folding at predicted “hinge” 

regions and that microfibril elasticity is 

dependent on a conformation-dependent 

maturation to the ~one-third staggered 

arrangement. 

Armed with this new structural information, 

we are now able to test our model by 

analysing the predicted hinge regions 

biochemically and through high-resolution 

structure determination. We are also 

defining higher-order microfibril packing 

arrangements by examining the biophysical 

properties of microfibril bundles and by 

studying microfibril interactions and crosslinks 

within bundles. Insights into the 

crucial role of microfibrils in elastic fibre 

formation are emerging from analysis of 

interactions with tropoelastin and elastinmicrofibril 

interface proteins. 

Clair Baldock and Cay Kielty 

page 19

Genetic Control of Tissue 

Structure and Function 

MICHAEL D BRIGGS BSc PhD 

ARC Research Fellow 

MICHAEL E GRANT BScTech 

D Phil 

Pro-Vice-Chancellor and 

Professor of Medical Biochemistry 

Molecular genetics and 

cell-matrix pathology of human 

monogenetic bone diseases 


Chapman,K.L., G.R.Mortier, K.Chapman, 

J.Loughlin, M.E.Grant, and M.D.Briggs. 2001. 

Mutations in the region encoding the von 

Willebrand factor A domain of matrilin-3 are 

associated with multiple epiphyseal dysplasia. 

Nat. Genet. 28:393-396. 

Holden,P., R.S.Meadows, K.L.Chapman, 

M.E.Grant, K.E.Kadler, and M.D.Briggs. 2001. 

Cartilage oligomeric matrix protein interacts with 

type IX collagen, and disruptions to these 

interactions identify a pathogenetic mechanism in 

a bone dysplasia family. J. Biol. Chem. 

276:6046-6055. 

Briggs,M.D. and Chapman, K.L. 2002. 

Pseudoachondroplasia and multiple epiphyseal 

dysplasia: mutation review, molecular interactions 

and genotype-phenotype correlation. Human 

Mutation. in press. 

Co-workers: 

Helen Attisha BSc 

Nuffield Foundation Student 

Kathryn Chapman BSc PhD 

ARC Research Associate 

Gail Skinner BSc 

European Commission Research 

Associate 

The skeletal dysplasias 

are an extremely 

diverse and complex 

group of genetic 

disorders, which primarily affect 

the development of the osseous 

skeleton. There are over 200 unique 

and well-characterised phenotypes, which range in severity from 

relatively mild to severe and lethal forms. Many of these 

individual phenotypes have been grouped into ‘bone dysplasia 

families’, on the basis of a similar clinical or radiographic 

presentation. One such family is the multiple epiphyseal 

dysplasia (MED) and pseudoachondroplasia (PSACH) group of 

diseases. 

PSACH appears to result almost exclusively from mutations in 

the gene encoding cartilage oligomeric matrix protein (COMP). 

Some forms of MED are allelic with PSACH, but MED shows 

considerable genetic heterogeneity and can also result from 

mutations in the genes encoding the α1, α2 and α3 chains of 

type IX collagen, COL9A1 (EDM6), COL9A2 (EDM2) and 

COL9A3 (EDM3), respectively. Furthermore, we have recently 

identified mutations in the gene encoding matrilin-3 (MATN3), a 

member of the matrilin family of extracellular oligomeric matrix 

proteins, which cause a distinctive mild form of MED (EDM5). 

The non-allelic genetic heterogeneity of the MED disease 

spectrum can best be explained by the observation that the 

protein products encoded by the COMP, type IX collagen and 

matrilin-3 genes interact to form large macromolecular 

assemblies in the cartilage ECM. Disruptions to these 

interactions are likely to have a fundamental effect on the 

development and homeostasis of the cartilage ECM and 

ultimately result in phenotypes within the PSACH-MED disease 

spectrum. 

We are using a multidisciplinary approach to study the molecular genetics and cell-matrix pathology 

of the PSACH and MED disease spectrum. Particular emphasis is being placed on understanding 

the non-allelic genetic heterogeneity of MED resulting from the disruption of supramolecular 

assemblies and the effect of specific disease causing mutations on the structure and function of the 

relevant gene products. Furthermore the cell, matrix and tissue pathology of matrilin-3 defects will 

be studied through the generation and analysis of an engineered mouse model. 

page 20

RAYMOND BOOT- 

HANDFORD BSc PhD 

Reader in Biochemistry and 

Molecular Biology 


Marks,D.S., C.A.Gregory, G.A.Wallis,A.Brass, 

K.E.Kadler, and R.P.Boot-Handford. 1999. 

Metaphyseal chondrodysplasia type Schmid 

mutations are predicted to occur in two distinct 

three-dimensional clusters within type X collagen 

NC1 domains that retain the ability to trimerize. 

J. Biol. Chem. 274:3632-3641. 

Fowler,S.J., S.Jose, X.Zhang, R.Deutzmann, 

M.P.Sarras, Jr., and R.P.Boot-Handford. 2000. 

Characterization of hydra type IV collagen.Type 

IV collagen is essential for head regeneration and 

its expression is up-regulated upon exposure to 

glucose. J. Biol. Chem. 275:39589-39599. 

Green,H.,A.E.Canfield, M.C.Hillarby, 

M.E.Grant, R.P.Boot-Handford,A.J.Freemont, and 

G.A.Wallis. 2000.The ribosomal protein QM is 

expressed differentially during vertebrate 

endochondral bone development. J. Bone Miner. 

Res. 15:1066-1075. 

Co-workers: 

Marianne Ellin BSc 

ARC Technician 

Darren Hitchen BSc 


Majid Shahbazi BSc PhD 


Biology, pathology and evolution 

of extracellular matrices 

The research in this laboratory covers two major areas: bone development and the 

pathogenesis of osteoarthritis, and invertebrate models for studying the pathology and evolution 

of extracellular matrix. 

Bone development and osteoarthritis. Much of the human skeleton is originally laid down during 

development as a cartilaginous template. The chondrocytes manufacturing this template are 

destined to progress through a highly coordinated differentiation pathway in the growth plate 

(see figure below) involving proliferation, maturation, hypertrophy (enlargement or swelling) and 

apoptosis. Cells brought to the location by vascular invasion erode the base of the growth plate 

and the matrix is converted from cartilage (type II collagen-rich) to bone (type I collagen-rich). 

The only chondrocytes not consumed in this process, known as endochondral ossification, are 

those that form the articular cartilage. These chondrocytes maintain the articular cartilage 

throughout life. However, in osteoarthritis, a disease characterised by a destruction of the 

articular cartilage matrix, the phenotype of articular chondrocytes is altered and they appear to 

de-differentiate and attempt to reinitiate the pattern of differentiation seen in the growth plate. 

We are using a number of different approaches, including gene targeting in vivo and cell culture 

models, to investigate the fundamental differences between growth plate and articular 

chondrocytes and determine whether the altered phenotype of these cells is a key factor in the 

pathogenesis of osteoarthritis. 

Invertebrate models. Hydra vulgaris is an evolutionarily ancient, simple, fresh water invertebrate 

composed of two epithelial cell sheets (ectoderm and endoderm) separated by a basement 

membrane-like matrix known as the mesoglea. Our interest in this organism stems from the fact 

that exposure to glucose leads to a thickening of the mesoglea in a fashion similar to that seen 

in basement membranes during diabetic microangiopathy – the secondary long term 

complication of diabetes which causes blindness and kidney failure. The glucose-induced 

mesoglea thickening in Hydra takes a few days whereas in diabetic patients, basement 

membrane thickening takes months to years. We have cloned the basement membrane (type 

IV) collagen gene from Hydra, as well as several other collagens that appear to contribute to the 

function of mesoglea, and shown it to be highly conserved in comparison to mammals. In 

addition, type IV collagen gene expression is upregulated within 48 hours of exposure of the 

Hydra to glucose. One aim is now to examine the mechanism by which glucose directly induces 

increases in type IV collagen synthesis in this relatively simple in vivo system since this may 

provide strong indications as to the aetiology of basement membrane thickening in diabetes. 

page 21

ULRIKE MAYER 

Dipl Chemistry PhD 

Lecturer in Dental Genetics 


Cohn,R.D., U.Mayer, G.Saher, R.Herrmann, 

.A.van der Flier,A.Sonnenberg, L.Sorokin, and 

T.Voit. 1999. Secondary reduction of alpha7B 

integrin in laminin alpha2 deficient congenital 

muscular dystrophy supports an additional 

transmembrane link in skeletal muscle. J. Neurol. 

Sci. 163:140-152. 

Miosge,N., C.Klenczar, R.Herken, M.Willem, and 

U.Mayer. 1999. Organization of the myotendinous 

junction is dependent on the presence of 

alpha7beta1 integrin. Lab Invest 79:1591- 

1599. 

Werner,A., M.Willem, L.L.Jones, G.W.Kreutzberg, 

U.Mayer, and G.Raivich. 2000. Impaired axonal 

regeneration in alpha7 integrin-deficient mice.J. 

Neurosci. 20:1822-1830. 

Co-workers: 

Melanie Klein MTA 


Peter Latham BSc PhD 


Synnva Ullensvang BSc 


Function of the laminin-nidogen-1 

interaction and the laminin-binding 

integrin α7β1 

The work of our group mainly focuses on the analysis of the basement membrane proteins 

laminin and nidogen in vivo and the receptor-mediated interaction of basement membranes 

with cells by gene targeting approaches. The combination of the analysis of the 

supramolecular organisation of basement membranes and the function of their cellular 

interactions will lead to a better understanding of a variety of biological processes. 

To study the potential involvement of the laminin-binding integrin α7β1 during myogenesis, and 

its role in muscle integrity and function, we have generated a null allele of the α7 gene in the 

germline of mice. Mice homozygous for the mutation are viable and fertile but develop a 

muscular dystrophy. We observed histopathological changes that strongly indicate an 

impairment of function of the myotendinous junctions. These findings demonstrate that α7β1 

integrin represents an indispensable linkage between muscle fibers and the extracellular matrix 

which is independent of the well characterised interaction of the cytoskeleton with the muscle 

basement membrane mediated by the dystrophin-dystroglycan complex. Ongoing studies 

include the elucidation of compensatory mechanisms by other integrin α subunits and the 

structural and functional relationship of the dystrophin/sarcoglycan complex with α7β1 integrin 

during myogenesis and muscle maturation. 

We are also studying the role of the laminin-nidogen-1 interaction in basement membrane 

assembly and function during embryogenesis and organ development. We have constructed a 

targeting vector for homologous recombination in embryonic stem cells in which the nidogen-1- 

binding module γ1III4 was deleted. This approach allows the study of a specialised interaction 

between two proteins without interfering with other biological functions known to be mediated 

by laminins. Mice homozygous for the deletion live to birth, but die soon after due to renal 

agenesis and impaired lung development. Future experiments include the generation of mice 

carrying subtle point mutations through which the laminin-nidogen-1 interaction is only 

weakened but not abolished. 

page 22

GILLIAN A.WALLIS BSc PhD 

Senior Lecturer in Medicine 


Roby,P., S.Eyre, J.Worthington, R.Ramesar, 

H.Cilliers, P.Beighton, M.Grant, G.Wallis. 1999. 

Autosomal dominant (Beukes) premature 

degenerative osteoarthropathy of the hip joint 

maps to an 11cM region on chromosome 4q35. 

Am J Hum Genet 64:904-908. 

White,A. and G.Wallis. 2001. Endochondral 

ossification:A delicate balance between growth 

and mineralisation. Current Biol. 11:R589-91. 

Newman,B., L.I.Gigout, L.Sudre, M.E.Grant and 

G.A.Wallis. 2001. Coordinated expression of 

matrix Gla protein is required during 

endochondral ossification for chondrocyte survival. 

J. Cell Biol. 154:659-666. 

Co-workers: 

Richard Aspinwall BSc MSc 

ARC Research Assistant 

Marianne Ellin BSc 

ARC Technician 

Emma Gillaspy BSc 

University of Manchester Student 

Carolyn Greig BSc 


Majid Shahbazi BSc PhD 


Kristian Spreckley BSc 


Laure Sudre BSc 

University Of Manchester Student 

The aetiology and pathogenesis of osteoarthritis 

Osteoarthritis (OA) is the most common form of human joint disease and a leading cause of 

pain and disability, particularly of the elderly. It is a heterogeneous condition in terms of 

cause, clinical course and severity. Furthermore, the pathogenesis is complex, involving 

both degradative and reparative processes of the articular cartilage and subchondral bone. 

We are approaching the study of OA in two ways. Firstly, we are investigating genetic 

susceptibility to the condition, and secondly, we are examining the processes whereby new 

bone is laid down in the OA joint. 

Epidemiological studies of OA have demonstrated that generalised nodal osteoarthritis 

(GNOA) has a strong genetic component. For our studies of genetic susceptibility to 

GNOA, we have obtained DNA from members of over 200 families that contain at least one 

affected sibling pair. We are genotyping these DNA samples using microsatellite markers 

that span the genome and analysing the data using non-parametric linkage analysis 

methods. We are using a candidate gene approach to investigate promising loci. 

During normal development, the endochondral ossification process is responsible for the 

conversion of the cartilaginous embryonic skeleton to bone. Recent studies have 

demonstrated that aspects of this process are reactivated in OA. We have identified 

several genes that are upregulated in cartilage both from persons with OA and in a naturally 

occurring form of OA in guinea pigs. We are investigating the function of these genes using 

a cell culture system that mimics endochondral ossification in vitro. We are developing 

methods for the transduction of genes that regulate the endochondral ossification process 

into chondrocytes in cartilage for future use in gene therapy for OA. 

The identification of genes important in the aetiology and progression of OA should allow us 

to identify persons most at risk for the condition and to develop strategies for its treatment. 

page 23

ANN E CANFIELD BSc PhD 

Senior Lecturer in Medicine 


Mantell,D.J., P.E.Owens, N.J.Bundred, E.B.Mawer 

and A.E.Canfield. 2000. 1 alpha,25- 

dihydroxyvitamin D(3) inhibits angiogenesis in 

vitro and in vivo. Circ. Res. 87:214-220. 

Canfield,A.E., M.J.Doherty,V.Kelly, B.Newman, 

C.Farrington, M.E.Grant and R.P.Boot-Handford. 

2000. Matrix Gla protein is differentially 

expressed during the deposition of a calcified 

matrix by vascular pericytes. FEBS Lett. 

487:267-271. 

Canfield,A.E., C.Farrington, M.D.Dziobon, 

R.P.Boot-Handford,A.M.Heagerty, S.N.Kumar 

and I.S.D.Roberts. 2002.The involvement of 

matrix glycoproteins in vascular calcification and 

fibrosis: an immunohistochemical study. J. Path. 

196: 228-234 

Co-workers: 

Yvonne Alexander BSc PhD 

Visiting Lecturer 

Georgina Collett BSc PhD 


Catherine Griffin-Jones HND 

EU-Technician 

Karen Howson BSc 

MRC Student 

Kirsty Ratcliffe BSc PhD 


Associate 

Claire Rock BSc PhD 

EU-Research Associate 

Neill Turner BSc 


Assistant 

Molecular and cellular mechanisms underpinning 

angiogenesis and vascular calcification 

Angiogenesis is the formation of new blood vessels from an existing vascular bed. It is 

of fundamental importance in many physiological and pathological conditions, including 

embryonic development, wound healing, atherosclerosis, diabetic retinopathy, psoriasis 

and tumour growth and metastasis. Angiogenesis is a complex process involving 

changes in endothelial cell phenotype, extracellular matrix remodelling and stabilisation 

of the newly formed blood vessels. We are currently employing a multidisciplinary 

approach: (i) to determine the contributions of specific proteins in mediating the 

response of endothelial cells to angiogenic factors during the early stages of 

angiogenesis, and (ii) to define the roles of specific angiogenic factors and matrix 

proteins in regulating endothelial cell-pericyte interactions that are crucial for vessel 

stabilisation. We aim to translate this work into the development of novel antiangiogenic 

and pro-angiogenic strategies for the treatment of diseases characterised by 

abnormal vascularisation. 

Calcification is associated with advanced complicated atherosclerosis in large arteries, 

but may also occur in smaller vessels where it results in ischemic tissue necrosis. We 

have shown that vascular pericytes can differentiate into osteoblast-like cells in vitro and 

in vivo, and that they can deposit a calcified matrix resembling that found in calcified 

atherosclerotic plaques. These results strongly suggest that pericytes may mediate, at 

least in part, vascular calcification. We are currently using molecular, cellular and 

biochemical approaches to elucidate the mechanisms of pericyte differentiation. This 

work is likely to provide important insights into the pathogenesis of vascular calcification. 

page 24

Localisation of MGP in calcified 

arteries.MGP is not detected 

in non-calcified vessels. 


Vascular Calcification 

Vascular calcification, a common 

complication of atherosclerosis and 

diabetes, leads to an increased risk of 

plaque rupture, myocardial infarction, limb 

amputation and morbidity. Interestingly, 

detailed studies of vascular calcification 

have shown it to be a highly complex 

process with many similarities to bone 

formation (osteogenesis). For example, 

proteins known to be involved in the 

controlled calcification that occurs during 

osteogenesis have also been identified in 

calcified atherosclerotic lesions. These 

proteins include bone morphogenetic 

protein-2 (BMP-2), BMP-6, osteopontin, 

osteocalcin, bone sialoprotein and matrix 

Gla protein. We have now shown that 

osteopontin and matrix Gla protein are also 

localised to sites of calcification in 

subcutaneous and dermal arteries, arterioles 

and microvessels in patients with calcific 

uraemic arteriolopathy (calciphylaxis), 

suggesting that calcification in these different 

sized vessels may actually occur by a 

common mechanism. 

The overarching aim of the research in my 

laboratory is to develop ways to control 

vascular calcification, and in recent studies 

we have aimed to elucidate how vascular 

calcification is regulated at the cellular and 

molecular level. Our findings have 

demonstrated that the deposition of mineral 

in arteries may be mediated by a subpopulation 

of smooth muscle cells and/or 

pericytes present in the vessel wall. These 

cells can differentiate into ‘osteoblast-like’ 

cells in vivo and in vitro, and deposit a 

calcified matrix resembling that found in 

calcified atherosclerotic plaques. To gain a 

better understanding of how this process is 

regulated at the molecular level, we have 

used subtractive hybridisation to identify 

genes that are either up-regulated or downregulated 

as pericytes undergo osteogenic 

differentiation and deposit a calcified matrix. 

Using this approach we have shown that 

genes that are implicated in the pathological 

calcification of arteries, namely matrix Gla 

protein, Axl receptor tyrosine kinase and 

HtrA1 serine protease are all differentially 

expressed during pericyte differentiation. 

Functional analyses of these genes using in 

vitro models developed in my laboratory 

have recently demonstrated that Axl, matrix 

Gla protein and HtrA1 play crucial, but 

distinct, roles in vascular calcification. The 

activation of Axl by its ligand (Gas6) 

appears to be required for maintaining 

these cells in their vascular phenotype. 

Down-regulation of Axl, or perturbation of 

Axl-Gas6 interactions, enhances the 

deposition of a calcified matrix by these 

cells. In contrast, matrix Gla protein appears 

to play a dual role in vascular cells, initially 

modulating cell differentiation and 

subsequently controlling matrix calcification. 

Interestingly, recent studies suggest that 

HtrA1 may regulate calcification by 

controlling the degradation of specific matrix 

proteins. We are currently conducting a 

detailed investigation of the mechanism(s) 

by which each of these proteins regulates 

calcification. 

In time, this integrated programme of 

research will provide a better understanding 

of the molecular events regulating 

physiological and pathological calcification, 

and may identify potential targets for the 

therapeutic manipulation of this event. 

Ann Canfield 

page 25

Facilities in the Centre 

The work in the Centre covers a broad span of activities 

encompassing approaches from pure population genetics, 

through to structural and functional studies of individual 

proteins and their assemblies. This breadth enables a rich 

interplay of interdisciplinary contacts but also requires an 

extensive infrastructure. The formation of the Centre was 

coupled to the establishment of a common core of 

equipment facilities. Our philosophy has been that wet 

laboratory space for people is precious and that where it is 

practical, equipment and other facilities are shared. This 

approach has greatly benefited new scientists coming to 

the Centre in rapidly establishing their laboratories and 

giving them access to state-of-the-art equipment. 

Biomolecular analysis 

The Centre has an extensive range of shared equipment 

for the purification, characterisation and study of the 

interactions of biomolecules. These multi-user facilities are 

housed in two fully integrated laboratories. 

The first of these laboratories is focused primarily on the 

isolation, identification and quantitation of proteins and 

glycoproteins. This laboratory is equipped with a range of 

chromatography workstations (standard, micro and 

capillary LC) used either for protein purification or as inlet 

devices for three of our four LC-mass spectrometers (see 

below). A liquid handling workstation enables automated 

multi-dimensional chromatographic separations. The mass 

spectrometers (a Tof-Spec E MALDI-TOF mass 

spectrometer, an LC-TOF mass spectrometer, a Quattro 

electrospray quadrapole mass spectrometer and a hybrid 

quadrapole-TOF mass spectrometer, all supplied by 

Micromass) are used for protein/glycoprotein identification, 

analysis of post-translational modifications, quantitation 

and accurate mass measurements. An automated N- 

terminal protein sequencer (Applied Biosystems) and a 

fluorescence spectrometer and CD spectrapolarimeter 

(Jasco) are also present in this laboratory. 

The second laboratory is focused on the physical analysis 

of biomolecules in solution and the investigation of their 

interactions. An extensive array of modern facilities is 

available for physical analysis of proteins, glycoproteins 

and polysaccharides, including a Beckman Optima XL-A 

analytical ultracentrifuge, a Dawn multi-angle light 

scattering photometer, a Malvern 4700 photon correlation 

spectrometer and a Viscotek protein analyser. 

Furthermore, this laboratory houses ultrasensitive 

calorimetry equipment (Microcal ITC and DSC) for the 

analysis of molecular transitions and the determination of 

binding constants, stoichiometries and reaction energies 

for interactions between biomolecules and their ligands. 

Electron microscopy 

The research of several groups in the Centre is dependent 

on our electron microscopy facility which contains a new 

Wellcome Trust-funded cryo-transmission electron 

microscope (Tecnai12 with STEM facility). Ancillary 

equipment is also available for freeze-fracture 

(Cressington), high pressure freezing (EM-Pact), freeze 

substitution as well as cryo-plunge equipment for use in 

cryo-EM. The Centre also has a dedicated image analysis 

suite that is used in automated electron tomography (AET) 

studies of extracellular matrix assemblies. 

Light Microscopy 

A major research focus within the Centre is live-cellimaging 

with a particular requirement for multicolour 

fluorescence. The Centre has an established 

microinjection facility that is also equipped with an 

environmentally controlled chamber, epi-fluorescence and 

software for time-lapse imaging. 

A “next generation” confocal microscopy facility is being 

established. These microscopes have the ability to 

separate spectral signatures and can optically section 

living cells with minimal damage using a variety of 

fluorescent probes including pairs of GFP variants. The x, 

y, z motorised stage enables accurate point visiting during 

time-lapse experiments. The system will have the 

capability for FRET, FRAP and FLIM studies. 

X-ray crystallography 

A new, state-of-the-art X-ray diffraction facility has been 

funded by the Wellcome Trust as part of the Centre 

renewal. This facility will consist of controlled-temperature 

crystallisation rooms, an X-ray diffraction generator 

equipped with high-brilliance optics, an area detector 

system, and a cryo-cooling system for diffracting frozen 

crystals. The facility will also include computational 

resources for data processing and analysis, 

crystallographic model building and refinement, and 

structural analysis. 

A fermentation suite with facilities for the bulk production of 

proteins from 5 – 20 L of media is also available. Five 

separate fermenters can accommodate bacterial, yeast 

and mammalian cultures. 

page 26

Research and Training 

Opportunities 

The Wellcome Trust Centre offers excellent training in all aspects of the biology of the 

extracellular matrix at all career levels. Staff in the Centre have an impressive track 

record in helping young researchers to develop independent careers, and they also 

provide opportunities for science graduates to conduct postgraduate and postdoctoral 

research. In addition, opportunities are available to help young clinicians gain 

appropriate support and undertake PhD or MD training. Each year opportunities exist 

to apply for: 

Senior Research Fellowships 

Career Development Fellowships 

Post-doctoral research positions 

Clinical Research and Research Training Fellowships (medical & veterinary) 

Postgraduate studentships 

Undergraduate Bursaries 

Pre-University School Bursaries 

Visiting (sabbatical) Fellowships 

Training for higher degrees in under the aegis of the School of Biological Sciences. 

The PhD and MPhil research training offered by the School is based on supportive 

supervision in an excellent research environment, a personal development 

programme for each student, and a structured Graduate Training Programme. The 

School also has a career development programme for Postdoctoral Research 

Associates and Fellows. The excellence and innovation of the postgraduate 

programme is recognised nationally and reflected in the annual award, by the 

Wellcome Trust, of five 4-year PhD studentships. Each year in the School, all research 

students participate in a two-day Graduate Symposium. In addition, students and 

postdoctoral researchers contribute to a weekly seminar programme, and are actively 

encouraged to present their results at national and international meetings. 

page 25 

page 27

Staff Profiles - Principal Investigators 

CLAIR BALDOCK 

MARTIN BARON 

JORDI BELLA 

1994 BSc Molecular Biophysics, 

University of Leeds. 

1997 PhD University of Sheffield. 

1998 Royal Society Study visit, 

University of Auckland, 

New Zealand. 

1998 Post-doctoral Research Associate, 

University of Manchester. 

2001 Royal Society Olga Kennard 

Research Fellow, 


1986 BA University of Oxford 

1990 DPhil University of Oxford 

1990 Post-doctoral Fellow, 

University of Oxford 

1991 Post-doctoral Fellow, 

Yale University 

1994 Lecturer, School of 

Biological Sciences, 

University of Manchester 

1984 BSc Chemistry, Universitat de 

Barcelona, Spain 

1991 PhD Chemical Sciences, 

Universitat Politècnica de 

Catalunya, Spain 

1991 Post-doctoral Fellow, Rutgers 

University, Piscataway, NJ, USA 


Purdue University, West Lafayette, 

IN, USA 

1999 Lecturer, School of Biological 

Sciences, University of Manchester 

PAUL BISHOP 

MIKE BRIGGS 

RAY BOOT-HANDFORD 

1980 B Med Sci University of 

Nottingham 

1983 BM BS University of Nottingham 

1988 FRCS London 

1991 Wellcome Trust Vision Research 

Training Fellow 

1993 PhD University of Manchester 

1994 Wellcome Trust Clinician 

Scientist Fellow 

1998 Honorary Consultant 

Ophthalmologist 

1999 Wellcome Trust Senior Research 

Fellow in Clinical Science 

1989 BSc Liverpool Polytechnic 

1993 PhD MRC Clinical Research 

Centre, Harrow 

1992 Post-doctoral Research Fellow, 

Cedars Sinai Medical Center, 

Los Angeles 

1996 ARC Research Fellow, 


2001 Reader in Biochemistry and 


1976 BSc University College of Wales, Cardiff 

1980 PhD University of London 

1981 Post-doctoral Fellow, University of 

Manchester 

1985 Post-doctoral Research Fellow, UMDNJ- 

Rutgers Medical School, Piscataway, NJ 

1987 RNIB Research Fellow, 


1989 School of Biological Sciences, 


1998 Wellcome Trust Research Leave 

Fellowship 

2001 Reader in Biochemistry and 


KEITH BRENNAN 

NEIL BULLEID 

ANN CANFIELD 

1994 BA University of Cambridge 

1998 PhD University of Cambridge 

1997 Wellcome Trust Prize Fellow at the 

University of Cambridge 

1999 Post-doctoral Research Fellow at 

the Strang Cancer Research 

Laboratories, New York, USA and 

Weill Medical College of Cornell 

University, New York, USA 

2001 School of Biological Sciences, 


2002 Wellcome Trust Research Career 

Development Fellow, 


ANDREW GILMORE 

1982 BSc University of Liverpool 

1985 PhD Glasgow College 


University of Kent 

1989 Visiting Post-doctoral Fellow, 

Howard Hughes Medical Institute, 

Dallas, Texas 

1990 Royal Society University Research 

Fellow,University of Manchester 

2000 Professor of Biochemistry, 

Univesity of Manchester 

MIKE GRANT 

1980 BSc University of Manchester 



Christie Hospital, NHS Trust, 

Manchester 

1993 Wellcome Trust Post-doctoral 

Research Fellow, University of 

Manchester 

1996 Lecturer, Dept of Medicine, 


2000 Senior Lecturer, Dept of Medicine, 


TIM HARDINGHAM 

1989 BA, Oxford University 

1993 PhD, University of Leicester 

1994 EMBO Post Doctoral Fellow, 

University of North Carolina 

1997 Postdoctoral Research Associate, 


2000 Wellcome Trust Research Career 



page 28 

1962 BSc Tech UMIST 

1966 DPhil Oxford University 

1966 Dept Medical Biochemistry 


1970 Post-doctoral Fellow 

University of Pennsylvania 

1972 University of Manchester 

1995 Chairman, Wellcome Trust 

Centre 

for Cell-Matrix Research, 


2000 Fellow of Academy of Medical 

Sciences 

2001 Pro-Vice-Chancellor, and 

Professor of Medical 

Biochemistry, 


1965 BSc University of Bristol 

1968 PhD University of Bristol 

1984 DSc University of Bristol 

1968 Kennedy Institute of Rheumatology 

1976 MRC Travelling Fellow, NIDR 

Bethesda, USA 

1995 Professor of Biochemistry, School 

of Biological Sciences, University of 

Manchester, (Wellcome Trust 

University Award) 

2001 Director, UK Centre for 

Tissue Engineering, 

University of Manchester

MARTIN HUMPHRIES 

KARL KADLER 

RICHARD KAMMERER 

1980 BSc University of Manchester 


1983 National Cancer Institute, NIH, USA 

and Howard University Cancer 

Center, Washington, D.C. USA 


Fellow University of Manchester 

1995 Wellcome Trust Principal Research 

Fellow and Professor of Biochemistry 


2000 Fellow of Academy of 

Medical Sciences 

2000 Director, Wellcome Trust Centre for 

Cell-Matrix Research, 


1980 BSc University of Salford 


1984 Post-doctoral Research Fellow 

UMDNJ-Rutgers Medical School, 

NJ USA 

1986 Assistant Professor, Thomas 

Jefferson University, Philadelphia, 

PA USA 


Fellow University of Manchester 

2000 Professor of Biochemistry, 


1992 Dipl Phil Biochemistry, Biozentrum, 

University of Basel, Switzerland 

1996 PhD Biochemistry, Biozentrum, 

University of Basel, Switzerland 


Biozentrum, University of Basel, 

Switzerland 

2000 Habilitation, University of Basel, 

Switzerland 

2000 Wellcome Trust Career 



CAY KIELTY 

ULRIKE MAYER 

JOHN SHEEHAN 

1978 BSc Kings College, London 

1981 PhD University College, London 



1990 Wellcome Trust Post-doctoral 

Research Fellow 

1993 MRC Senior Research Fellow 

2000 Professor of Medical Biochemistry, 


2001 Fellow of Academy of 

Medical Sciences 

1984 Dipl. Chemistry, Albert-Ludwig 

University Freiburg, Germany 

1988 PhD Biochemistry, 

Max-Planck-Institute for Biochemistry, 

Martinsried, Germany 


Max-Planck-Institute for Biochemistry, 

Martinsried, Germany 


Max-Planck-Group for Rheumatology 

and Immunology, Erlangen, Germany 

1994 Research Fellow, Max-Planck-Institute 

for Biochemistry, Martinsried, 

Germany 

2000 Lecturer in Dental Genetics, 


1968 BSc Physics, University of Leicester 

1970 MSc Materials Science, 

University of Bristol 

1973 PhD Biophysics, University of Bristol 


University of Bristol 


University of Lancaster 

1979 Research Fellow, 

University of Lund, Sweden 


University of Lancaster 

1988 Wellcome Trust Senior Lecturer, 


1998 Reader in Physiological Biochemistry, 

School of Biological Sciences, 


ADRIAN SHUTTLEWORTH 

CHARLES STREULI 

DANNY TUCKWELL 

1964 BSc University of Liverpool 

1967 PhD University of Liverpool 


Harvard Medical School 


Northwestern University Medical 

School, Chicago 

1969 Lecturer, Dept of Medical 

Biochemistry, University of 

Manchester 

1996 Reader in Medical Biochemistry, 


1979 BA University of Cambridge 

1982 MA University of Cambridge 

1983 PhD University of Leicester 

1982 Post-doctoral Research Fellow, ICRF, 

London 


RPMS, London 

1987 Senior post-doctoral Scientist, 

Lawrence Berkeley Laboratory, 

Berkeley, USA 


Fellow, University of Manchester 

1986 BSc University of Bristol 

1990 PhD University of Nottingham 



1997 BBSRC Advanced Research Fellow 

GILLIAN WALLIS 

1982 BSc University of Cape Town, 

South Africa 

1985 PhD University of Cape Town, 

South Africa 

1985 Senior Research Associate, Dept 

of Human Genetics, UCT 


University of Washington, 

Seattle, USA 



1996 Lecturer Dept of Medicine, 


2000 Senior Lecturer, Dept of Medicine, 


page 29

Staff List January 2002 

DEVELOPMENT OFFICER 

Linda Green 

COMPUTER MANAGER 

Adam Huffman 

SECRETARY 

Carol McMurdo 

SENIOR EXPERIMENTAL 

OFFICER 

Dave Thornton 

TECHNICAL STAFF 

Susan Allan 

Janet Askari 

Stephanie Barton 

Jane Bradley 

Seema Chakravarthi 

Sue Craig 

Marianne Ellin 

Catherine Griffin Jones 

Darren Hitchen 

Marj Howard 

Emma Keevill 

Tracy Kilbride 

Melanie Klein 

Jane Kott 

Amanda Lomas 

Emma Lowe 

Roger Meadows 

Kieran Mellody 

Amanda Morgan 

Eileen Pinnington 

Janine Prince 

Synnva Ullensvang 

Rachel Watkins 

CLINICAL FELLOWS AND 

LECTURERS 

Yvonne Alexander 

Chris Brew 

Dave Murray 

Wang Jing 

POSTDOCTORAL RESEARCH 

ASSOCIATES 

Adetola Adesida 

Nasreen Akhtar 

Steve Ball 

Mark Bass 

Dan Bax 

Patrick Buckley 

Elizabeth Canty 

Kathryn Chapman 

Georgina Collett 

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Laure Garrique-Antar 

John Hindson 

Catherine Hoare 

Nigel Hodson 

David Holmes 

Karen Johnston 

Tom Jowitt 

Saiqa Khan 

Peter Latham 

Magali Le Goff 

David Lee 

Matthew Leighton 

Emma Marshman 

Anthea Messent 

Zohreh Mostafavi-Pour 

Paul Mould 

Alan Murdoch 

QingQiu Pu 

Gareth Pugh 

Pooli Ragesekariah 

Kirsty Ratcliffe 

Susan Richardson 

Claire Rock 

Matt Rock 

Majid Shahbazi 

Mike Sherratt 

Gail Skinner 

Stephen St. George Smith 

Toby Starborg 

Simon Stephan 

Mohammed Tasab 

Simon Tew 

Pengbo Wang 

Mark Warren 

Marian Wilkin 

RESEARCH ASSISTANTS 

Richard Aspinwall 

Craig Barley 

Tanja Benkert 

Carolyn Greig 

Jon Humphries 

Sara Kirkham 

Philip Macdonald 

Ngoc-sa Nguyen Huu 

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Neill Turner 

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Anthony Valentijn 

POSTGRADUATE STUDENTS 

Helen Attisha 

Sarah Bernard 

Claire Bithell 

Ann-Marie Carbery 

Claire Crouchley 

Linda Eyers 

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Hanane Gouizi 

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Vasiliki Petropoulou 

Suzannah Phillips 

Richard Rauchenberg 

Adam Shaw 

Laure Sudre 

Emlyn Symonds 

Mark Travis 

Matthew Wakefield 

Harriet Watkin 

Nadia Zouq 

page 30

PUBLICATIONS 2000 - 2001 

Addinall,S.G., P.S.Mayr, S.Doyle, 

J.K.Sheehan, P.G.Woodman and V.J.Allan. 

2001. Phosphorylation by cdc2-CyclinB1 

kinase releases cytoplasmic dynein from 

membranes. J. Biol. Chem. 

276:15939-15944. 

Alexandrescu,A.T., M.W.Maciejewski, 

M.A.Ruegg, J.Engel and R.A.Kammerer. 

2001. 1H, 13C and 15N backbone 

assignments for the C-terminal globular 

domain of agrin. J. Biomol. NMR 

20:295-296. 

Almond,A., A.Brass and J.K.Sheehan. 2000. 

Oligosaccharides as model systems for 

understanding water-biopolymer interaction: 

hydrated dynamics of a hyaluronan 

decamer. J. Phys. Chem. B. 

104:5634-5640. 

Almond,A. and J.K.Sheehan. 2000. 

Glycosaminoglycan conformation: do 

aqueous molecular dynamics simulations 

agree with x-ray fibre diffraction? 

Glycobiology 10:329-338. 

Anderton,L. and P.Bishop. 2001. Acute 

visual loss following thioridazine overdose. 

Am. J. Psychiatry. 158: 818. 

Aquilina,A., M.Korda, J.M.Bergelson, 

M.J.Humphries, R.Farndale and 

D.S.Tuckwell. 2002. A novel gain of function 

mutation in the integrin α2 VWFA domain; 

European Journal of Biochem. in press 

Ashworth,J.L., C.M.Kielty and D.McLeod. 

2000. Fibrillin and the eye. Br. J. 

Ophthalmol. 84:1312-1317. 

Aszódi,A., J.F. Bateman, E.Gustafsson, 

R.P.Boot-Handford and R.Fässler. 2000 

Mammalian skeletogenesis and extracellular 

matrix: What can we learn from knockout 

mice? Cell Structure and Function 

25:71-82. 

Baldock,C., A.J.Koster, U.Ziese, M.J.Rock, 

M.J.Sherratt, K.E.Kadler, C.A.Shuttleworth 

and C.M.Kielty. 2001. The supramolecular 

organization of fibrillin-rich microfibrils. J. 


Ball,S.G., C.Baldock, C.M.Kielty and 

C.A.Shuttleworth. 2001. The role of the C1 

and C2 A-domains in type VI collagen 

assembly. J. Biol. Chem. 276:7422-7430. 

Baron,M., V.O’Leary, D.A.Evans, M.Hicks 

and K.Hudson. 2000. Multiple roles of the 

Dcdc42 GTPase during wing development in 

Drosophila melanogaster. Mol. Gen. Genet. 

264:98-104. 

Bella,J. and H.M.Berman. 2000. Integrincollagen 

complex: a metal-glutamate 

handshake. Structure Fold. Des. 

8:R121-R126. 

Bella,J. and M.G.Rossmann. 2000. The 

dynamics of receptor recognition by human 

rhinoviruses: response. Trends Microbiol. 

8:254. 

Bella,J. and M.G.Rossmann. 2000. ICAM-1 

receptors and cold viruses. 

Pharm. Acta Helv. 74:291-297. 

Bishop,P.N. 2000. Structural 

macromolecules and supramolecular 

organisation of the vitreous gel. Prog. Retin. 

Eye Res. 19:323-344. 

Bishop,PN. 2000. Laser treatments in 

age-related macular degeneration. 

CE Optometry. 3:100-103. 

Bishop,PN., 2000. Matrix metalloproteinases 

and their natural inhibitors in fibrovascular 

membranes of proliferative diabetic 

retinopathy (editorial). Br. J. Ophthalmol. 

84:1087-1088. 

Biswas,S., F.L.Munier, J.Yardley, N.Hart- 

Holden, R.Perveen, P.Cousin, J.E.Sutphin, 

B.Noble, M.Batterbury, C.M.Kielty, 

A.Hackett, R.Bonshek, A.Ridgway, 

D.McLeod, V.C.Sheffield, E.M.Stone, 

D.F.Schorderet and G.C.M.Black. 2001. 

Missense mutations in COL8A2, the gene 

encoding the α 2 chain of type VIII collagen, 

cause two forms of corneal endothelial 

dystrophy. Hum. Mol. Genet. 

10: 2415-2423. 

Bos,K.J., D.F.Holmes, K.E.Kadler, 

D.McLeod, N.P.Morris and P.N.Bishop. 

2001. Axial structure of the heterotypic 

collagen fibrils of vitreous humour and 

cartilage. J. Mol. Biol. 306:1011-1022. 

Bos,K.J., D.F.Holmes, R.S.Meadows, 

K.E.Kadler, D.McLeod and P.N.Bishop. 

2001. Collagen fibril organisation in 

mammalian vitreous by freeze etch/rotary 

shadowing electron microscopy. Micron 

32:301-306. 

Bottomley,M.J., M.R.Batten, R.A.Lumb and 

N.J.Bulleid. 2001. Quality control in the 

endoplasmic reticulum. PDI mediates the 

ER retention of unassembled procollagen C- 

propeptides. Curr. Biol. 11:1114-1118. 

Boudko,S., S.Frank, R.A.Kammerer, 

J.Stetefeld, T.Schulthess, R.Landwehr, 

A.Lustig, H.P.Bächinger and J.Engel. 2002 

Nucleation and propagation of the collagen 

triple helix in single-chain and trimerized 

peptides: transition from 3rd to 1st order 

kinetics. J. Mol. Biol., in press. 

Briggs,M.D. 2000. Screening for mutations 

in cartilage ECM genes. Methods Mol. Biol. 

139:133-145. 

Briggs,M.D. and Chapman,K.L. 2002. 

Pseudoachondroplasia and multiple 

epiphyseal dysplasia: mutation review, 

molecular interactions and genotypephenotype 

correlation. Human Mutation. 

in press. 

Brookman,J.L., G.Mennim, A.P.Trinci, 

M.K.Theodorou and D.S.Tuckwell. 2000. 

Identification and characterization of 

anaerobic gut fungi using molecular 

methodologies based on ribosomal ITS1 

and 18S rRNA. Microbiology 146: 393-403. 

Bulleid,N.J., D.C.John and K.E.Kadler. 2000. 

Recombinant expression systems for the 

production of collagen. Biochem. Soc. 

Trans. 28:350-353. 

Burkhard,P., R.A.Kammerer, M.O.Steinmetz, 

G.P.Bourenkov and U.Aebi. 2000. The 

coiled-coil trigger site of the rod domain of 

cortexillin I unveils a distinct network of 

interhelical and intrahelical salt bridges. 

Structure Fold. Des. 8:223-230. 

Cabibbo,A., M.Pagani, M.Fabbri, M.Rocchi, 

M.R.Farmery, N.J.Bulleid and R.Sitia. 2000. 

ERO1-L, a human protein that favors 

disulfide bond formation in the endoplasmic 

reticulum. J. Biol. Chem. 275:4827-4833. 

Canfield,A.E., M.J.Doherty, V.Kelly, 

B.Newman, C.Farrington, M.E.Grant and 

R.P.Boot-Handford. 2000. Matrix Gla protein 

is differentially expressed during the 

deposition of a calcified matrix by vascular 

pericytes. FEBS Lett. 487:267-271. 

Canfield,A.E., M.J.Doherty and B.A.Ashton. 

2000. Osteogenic potential of vascular 

pericytes. In:Bone Engineering Eds. J.E. 

Davies. em squared incorporated, Toronto, 

Canada, pp143-151. 

Canfield,A.E., M.J.Doherty, A.C.Wood, 

C.Farrington, B.Ashton, N.Begum, B.Harvey, 

A.Poole, M.E.Grant and R.P.Boot-Handford. 

2000. Role of pericytes in vascular 

calcification: a review. Z. Kardiol. 

89 Suppl 2:20-27. 

Canfield,A.E., C.Farrington, M.D.Dziobon, 

R.P.Boot-Handford, A.M.Heagerty, 

S.N.Kumar and I.S.D.Roberts. 2002. The 

involvement of matrix glycoproteins in 

vascular calcification and fibrosis: an 

immunohistochemical study. J. Path. 

196: 228-234 

Chapman,K.L., G.R.Mortier, K.Chapman, 

J.Loughlin, M.E.Grant and M.D.Briggs. 

2001. Mutations in the region encoding the 

von Willebrand factor A domain of matrilin-3 

are associated with multiple epiphyseal 

dysplasia. Nature Genet. 28:393-396. 

Chaudhry,S.S., J.Gazzard, C.Baldock, 

J.Dixon, M.J.Rock, G.C.Skinner, K.P.Steel, 

C.M.Kielty and M.J.Dixon. 2001. Mutation of 

the gene encoding fibrillin-2 results in 

syndactyly in mice. Hum. Mol. Genet. 

10:835-843. 

Cheung,J.O., M.C.Hillarby, S.Ayad, 

J.A.Hoyland, C.J.Jones, J.Denton, 

J.T.Thomas, G.A.Wallis and M.E.Grant. 

2001. A novel cell culture model of 

chondrocyte differentiation during 

mammalian endochondral ossification. J. 

Bone Miner. Res. 16:309-318. 

page 31

Clark,K., P.Newham, L.Burrows, J.A.Askari 

and M.J.Humphries. 2000. Production of 

recombinant soluble human integrin α4β1. 

FEBS Lett. 471:182-186. 

Coe,A.P., J.A.Askari, A.D.Kline, 

M.K.Robinson, H.Kirby, P.E.Stephens and 

M.J.Humphries. 2001. Generation of a 

minimal α5β1 integrin-Fc fragment. J. Biol. 

Chem. 276:35854-35866. 

Costes,S., C.H.Streuli and M.H.Barcellos- 

Hoff. 2000. Quantitative image analysis of 

laminin immunoreactivity in skin basement 

membrane irradiated with 1 GeV/nucleon 

iron particles. Radiat. Res. 154:389-397. 

Day,A.J. and Sheehan,J.K. 2001. 

Hyaluronan: polysaccharide chaos to protein 

organisation. Current Opinion in Structural 

Biology. 11:617-622. 

Deutzmann,R., S.Fowler, X.Zhang, K.Boone, 

S.Dexter, R.P.Boot-Handford, R.Rachel and 

M.P.Sarras, Jr. 2000. Molecular, biochemical 

and functional analysis of a novel and 

developmentally important fibrillar collagen 

(Hcol-I) in hydra. Development 

127:4669-4680. 

Engel,J. and R.A.Kammerer. 2000. What are 

oligomerization domains good for? Matrix 

Biol. 19:283-288. 

Farmery,M.R., S.Allen, A.J.Allen and 

N.J.Bulleid. 2000. The role of ERp57 in 

disulfide bond formation during the 

assembly of major histocompatibility 

complex class I in a synchronized 

semipermeabilized cell translation system. J. 

Biol. Chem. 275:14933-14938. 

Farmery,M.R. and N.J.Bulleid. 2001. Major 

histocompatibility class I folding, assembly 

and degradation: a paradigm for two-stage 

quality control in the endoplasmic reticulum. 

Prog. Nucleic Acid Res. Mol. Biol. 

67:235-268. 

Farrington,C., A.M.Heagerty and 

A.E.Canfield. 2000. Thrombospondin 

expression by calcifying and non-calcifying 

vascular cells. In: Chemistry and Biology 

of Mineralised Tissues. Eds.Goldberg, M. 

Boskey, A & Robinson, C. American 

Academy of Orthopaedic Surgeons. 

pp 377-381. 

Firth,L. J.Manchester, J.A.Lorenzen, 

M.Baron and L.A.Perkins. 2000. 

Identification of genomic regions that 

interact with a viable allele of the Drosophila 

protein tyrosine phosphatase corkscrew. 

Genetics 156:733-748. 

Fosang,A.J., K.Last, H.Stanton, D.B.Weeks, 

I.K.Campbell, T.E.Hardingham and 

R.M.Hembry. 2000. Generation and novel 

distribution of matrix metalloproteinasederived 

aggrecan fragments in porcine 

cartilage explants. J. Biol. Chem. 

275:33027-33037. 

Fowler,S.J., S.Jose, X.Zhang, R.Deutzmann, 

M.P.Sarras, Jr. and R.P.Boot-Handford. 

2000. Characterization of hydra type IV 

collagen. Type IV collagen is essential for 

head regeneration and its expression is upregulated 

upon exposure to glucose. J. Biol. 

Chem. 275:39589-39599. 

Frank,S., R.A.Kammerer, S.Hellstern, 

S.Pegoraro, J.Stetefeld, A.Lustig, L.Moroder 

and J.Engel. 2000. Toward a high-resolution 

structure of phospholamban: design of 

soluble transmembrane domain mutants. 

Biochemistry 39:6825-6831. 

Frank,S., A.Lustig, T.Schulthess, J.Engel 

and R.A.Kammerer. 2000. A distinct sevenresidue 

trigger sequence is indispensable 

for proper coiled-coil formation of the human 

macrophage scavenger receptor 

oligomerization domain. J. Biol. Chem. 

275:11672-11677. 

Frank,S., R.A.Kammerer, D.Mechling, 

T.Schulthess, R.Landwehr, J.Bann, Y.Guo, 

A.Lustig, H.P.Bachinger and J.Engel. 2001. 

Stabilization of short collagen-like triple 

helices by protein engineering. J. Mol. Biol. 

308:1081-1089. 

Garrigue-Antar,L., C.Barker and K.E.Kadler. 

2001. Identification of amino acid residues in 

bone morphogenetic protein-1 important for 

procollagen C-proteinase activity. J. Biol. 

Chem. 276:26237-26242. 

Gilmore,A.P., A.D.Metcalfe, L.H.Romer and 

C.H.Streuli. 2000. Integrin-mediated survival 

signals regulate the apoptotic function of 

Bax through its conformation and subcellular 


Gilmore, A.P. and C.H.Streuli. 2002. 

Analysing apoptosis in cultured epithelial 

cells. Methods Mol. Biol. in press. 

Graham,H.K., D.F.Holmes, R.B.Watson and 

K.E.Kadler. 2000. Identification of collagen 

fibril fusion during vertebrate tendon 

morphogenesis. The process relies on 

unipolar fibrils and is regulated by collagenproteoglycan 

interaction. J. Mol. Biol. 

295:891-902. 

Grant,M.E. 2001. Basement Membranes: 

more matrix than membrane. Biological 

Sciences Review 13:36-39. 

Green,H., A.E.Canfield, M.C.Hillarby, 

M.E.Grant, R.P.Boot-Handford, 

A.J.Freemont and G.A.Wallis. 2000. The 

ribosomal protein QM is expressed 

differentially during vertebrate endochondral 

bone development. J. Bone Miner. Res. 

15:1066-1075. 

Gregory,C.A., B.Zabel, M.E.Grant, R.P.Boot- 

Handford and G.A.Wallis. 2000. Equal 

expression of type X collagen mRNA from 

mutant and wild type COL10A1 alleles in 

growth plate cartilage from a patient with 

metaphyseal chondrodysplasia type Schmid. 

J. Med. Genet. 37:627-629. 

Gribbon,P., B.C.Heng and T.E.Hardingham. 

2000. The analysis of intermolecular 

interactions in concentrated hyaluronan 

solutions suggest no evidence for chainchain 

association. Biochem. J. 

350: 329-335. 

Gribbon,P., B.C.Heng and T.E.Hardingham. 

2001. Novel confocal-FRAP analysis of 

carbohydrate-protein interactions within the 

extracellular matrix. Methods Mol. Biol. 

171:487-494. 

Guo,Y., R.A.Kammerer and J.Engel. 2000. 

The unusually stable coiled-coil domain of 

COMP exhibits cold and heat denaturation 

in 4-6 M guanidinium chloride. Biophys. 

Chem. 85:179-186. 

Hadjiloucas,I., A.P.Gilmore, N.J.Bundred and 

C.H.Streuli. 2001. Assessment of apoptosis 

in human breast tissue using an antibody 

against the active form of caspase 3: 

relation to histopathological characteristics. 

Brit. J. Cancer 85: 1522-1526. 

Hardingham,T.E. and P.Gribbon. 2000. 

Confocal-FRAP analysis of ECM molecular 

interactions. In: Extracellular Matrix 

Protocols. Eds. Streuli C., Grant M.E. 

Humana Press pp. 83-93. 

He,Y., V.D.Bowman, S.Mueller, C.M.Bator, 

J.Bella, X.Peng, T.S.Baker, E.Wimmer, 

R.J.Kuhn and M.G.Rossmann. 2000. 

Interaction of the poliovirus receptor with 

poliovirus. Proc. Natl. Acad. Sci. U. S. A 

97:79-84. 

Hendershot,L.M. and N.J.Bulleid. 2000. 

Protein-specific chaperones: the role of 

hsp47 begins to gel. Curr. Biol. 

10:R912-R915. 

Hicks,M.S., V.O’Leary, M.Wilkin, S.E.Bee, 

M.J.Humphries and M.Baron. 2001. 

DrhoGEF3 encodes a new Drosophila DH 

domain protein that exhibits a highly 

dynamic embryonic expression pattern. Dev. 

Genes Evol. 211:263-267. 

Hill,J., M.Lewis, P.Mills and C.M.Kielty. 2002. 

Effects of pulsed short-wave diathermy on 

human fibroblast proliferation. Arch. Phys. 

Med. Rehabilitation in press. 

Holden,P., R.S.Meadows, K.L.Chapman, 

M.E.Grant, K.E.Kadler and M.D.Briggs. 

2001. Cartilage oligomeric matrix protein 

interacts with type IX collagen and 

disruptions to these interactions identify a 

pathogenetic mechanism in a bone 

dysplasia family. J. Biol. Chem. 

276:6046-6055. 





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organization. Proc. Natl. Acad. Sci. U. S. A 

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Holmes,D.F., H.K.Graham, J.A.Trotter and 

K.E.Kadler. 2001. STEM/TEM studies of 

collagen fibril assembly. Micron. 

32:273-285. 

Humphries,J.D., J.A.Askari, X.P.Zhang, 

Y.Takada, M.J.Humphries and A.P.Mould. 

2000. Molecular basis of ligand recognition 

by integrin α5β1. II. Specificity of Arg-Gly- 

Asp binding is determined by Trp157 of the 

α subunit. J. Biol. Chem. 275:20337-20345. 

Humphries,M.J. 2000. Integrin structure. 

Biochem. Soc. Trans. 28:311-339. 

Humphries,M.J. 2000. Cell adhesion assays. 

Methods Mol. Biol. 139:279-285. 

Humphries,M.J. 2000. Integrin cell adhesion 

receptors and the concept of agonism. 

Trends Pharmacol. Sci. 21:29-32. 

Humphries,M.J. 2000. Genetics and the 

biological functions of integrin adhesion 

receptors. Matrix Biol. 19:189-190. 

Humphries,M.J. and R.C.Liddington. 2000. 

Molecular basis of integrin-dependent cell 

adhesion. In: Frontiers in Molecular 

Biology: Protein-Protein Recognition. Ed. 

Kleanthous, C., Oxford University Press, 

pp. 102-125. 

Humphries,M.J. and A.P.Mould. 2001. An 

anthropomorphic integrin. Science 

294:316-317. 

Humphries,M.J. 2001. Cell adhesion assays. 

Mol. Biotechnol. 18:57-61. 

Illidge,C., C.Kielty and A.Shuttleworth. 2001. 

Type VIII collagen: heterotrimeric chain 

association. Int. J. Biochem. Cell Biol. 

33:521-529. 

Jackson,T., W.Blakemore, J.W.Newman, 

N.J.Knowles, A.P.Mould, M.J.Humphries and 

A.M.King. 2000. Foot-and-mouth disease 

virus is a ligand for the high-affinity binding 

conformation of integrin α5β1: influence of 

the leucine residue within the RGDL motif on 

selectivity of integrin binding. J. Gen. Virol. 

81: 1383-1391. 

Kadler,K.E., D.F.Holmes, H.Graham and 

T.Starborg. 2000. Tip-mediated fusion 

involving unipolar collagen fibrils accounts 

for rapid fibril elongation, the occurrence of 

fibrillar branched networks in skin and the 

paucity of collagen fibril ends in vertebrates. 

Matrix Biol. 19:359-365. 

Kammerer,R.A., V.A.Jaravine, S.Frank, 

T.Schulthess, R.Landwehr, A.Lustig, 

C.Garcia-Echeverria, A.T.Alexandrescu, 

J.Engel and M.O.Steinmetz. 2001. An 

intrahelical salt bridge within the trigger site 

stabilizes the GCN4 leucine zipper. J. Biol. 

Chem. 276:13685-13688. 

Kielty,C.M., C.Baldock, M.J.Rock, 

J.L.Ashworth and C.A.Shuttleworth. 2002. 

Fibrillin: from microfibril assembly to 

biomechanical function. Philosoph. Trans. 

Royal Soc. B. in press. 

Kielty,C.M. and M.E.Grant. 2002. The 

collagen family: structure, assembly and 

organization in the extracellular matrix. In: 

Connective Tissue and its Heritable 

Diseases; Molecular, Genetic and Medical 

Aspects. Eds.Royce, P.M. and Steinmann, 

B., Wiley-Liss, New York. in press. 

Kielty,C.M. 2002. Fibrillin: biomechanical 

function of fibrillin-rich microfibrils. Chapter 

In: Elastomeric proteins Eds. Shewry,P. & 

Bailey,A.J., Royal Society Discussion 

Meeting, Cambridge University Press. 

in press. 

Kirkham,S., J.K.Sheehan, D.Knight, 

P.S.Richardson and D.J.Thornton. 2002 

Heterogeneity of airways mucus: variations 

in the amounts and glycoforms of the major 

oligomeric mucins MUC5AC and MUC5B. 

Biochem J. 361: 537-546. 

Klinowska,T.C. and C.H.Streuli. 2000. 

Analyzing cell-ECM interactions in adult 

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page 35

PRINCIPAL INVESTIGATORS 

Dr Clair Baldock 0161 275 5439 clair.baldock@man.ac.uk 

Dr Martin Baron 0161 275 5111 martin.baron@man.ac.uk 

Dr Jordi Bella 0161 275 5467 jordi.bella@man.ac.uk 

Dr Paul Bishop 0161 275 5755 paul.bishop@man.ac.uk 

Dr Ray Boot-Handford 0161 275 5097 ray.boot-handford@man.ac.uk 

Dr Keith Brennan 0161 275 1517 keith.brennan@man.ac.uk 

Dr Mike Briggs 0161 275 5642 mike.briggs@man.ac.uk 

Professor Neil Bulleid 0161 275 5103 neil.bulleid@man.ac.uk 

Dr Ann Canfield 0161 275 5066 ann.canfield@man.ac.uk 

Dr Andrew Gilmore 0161 275 3892 andrew.gilmore@man.ac.uk 

Professor Mike Grant 0161 275 5074 michael.grant@man.ac.uk 

Professor Tim Hardingham 0161 275 5511 timothy.e.hardingham@man.ac.uk 

Professor Martin Humphries 0161 275 5071 martin.humphries@man.ac.uk 

Professor Karl Kadler 0161 275 5086 karl.kadler@man.ac.uk 

Dr Richard Kammerer 0161 275 1504 richard.kammerer@man.ac.uk 

Professor Cay Kielty 0161 275 5739 cay.kielty@man.ac.uk 

Dr Ulrike Mayer 0161 275 5246 ulrike.mayer@man.ac.uk 

Dr John Sheehan 0161 275 5647 john.sheehan@man.ac.uk 

Dr Adrian Shuttleworth 0161 275 5079 charles.a.shuttleworth@man.ac.uk 

Dr Charles Streuli 0161 275 5626 charles.streuli@man.ac.uk 

Dr Danny Tuckwell 0161 275 7392 danny.tuckwell@man.ac.uk 

Dr Gillian Wallis 0161 275 5629 gillian.a.wallis@man.ac.uk 

GENERAL ENQUIRIES TO: 

Dr. Linda J. Green, Development Officer, 

Wellcome Trust Centre for Cell-Matrix 

Research, School of Biological Sciences, 

University of Manchester, 2.205 Stopford 

Building,Oxford Road, Manchester, M13 

9PT, UK. 

Tel: +44-(0)161 275 1516 

Fax: +44(0)161 275 1505 

Email: linda.j.green@man.ac.uk

Review and Prospectus 2002 - Wellcome Trust Centre For Cell ...

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