Book of Abstracts - Ruhr-Universität Bochum

OP-17
ISBOMC `10 5.7 – 9.7. 2010 Ruhr-Universität Bochum
Biofunctionalization of a Generic Collagenous Triple Helix
with the Integrin α2β1 Binding Site
Stephan Niland, a Christoph Westerhausen, b Thilo Bracht, a Alletta Schmidt-Hederich, a Désirée Freund, a
Stephan W. Schneider, c Matthias F. Schneider, d and Johannes A. Eble a
a Goethe University Frankfurt, University Hospital, Center for Molecular Medicine, Department of
Vascular Matrix Biology, Excellence Cluster Cardio-Pulmonary System, Theodor-Stern-Kai 7, 60590
Frankfurt/Main, Germany. b University of Augsburg, Faculty of Mathematics and Natural Sciences,
Department of Experimental Physics I, Universitaetsstrasse 1, 86135 Augsburg, Germany.
c Heidelberg University, Mannheim University Hospital, Department of Experimental Dermatology,
Theodor-Kutzer-Ufer 1-3, 68167 Mannheim, Germany. d Boston University, Department of
Mechanical Engineering, Biological Physics, 590 Commonwealth Avenue, MA 02215, Boston, USA.
E-mail: niland@med.uni-frankfurt.de
Integrin ���� is a major collagen-binding receptor and widely distributed on different tissues. It plays
essential roles in elementary cell functions such as adhesion morphology, migration, proliferation,
gene activation, and differentiation. Therefore, it regulates various (patho)physiological situations,
such as thrombosis, tumor infiltration, and metastasis. It plays an essential role in liver micrometastasis
formation. 1
Integrin ���� recognizes triple-helical collagens, whose quaternary structure consists of three lefthanded
polyproline-like chains supercoiled in a right-handed helix about a common axis, yielding a
characteristic triple helical coiled-coil, which provides the framework for the integrin ����
recognition motif (GFOGER)3. Due to these structural requirements, chemical imitation of triplehelical
���� integrin recognition site is still a demanding feat. The aim of this study was to generate a
recombinant mini-collagen with a single binding site for integrin ���� and to use this collagenmimetic
to characterize the role of integrin �����at both molecular and cellular level. Force
spectroscopy was used to determine at the molecular level the binding of integrin �����to its triplehelical
recognition motif (GFPGER)3. To define and disclose the role of integrin �����in cell biology,
an integrin ����-specific and agonistic recombinant mini-collagen was used, as well as the snake
venom-derived highly specific integrin ���� antagonist rhodocetin. 2
The strong binding of integrin �����to its ligand underlines its importance as cellular
mechanotransducer. On a substratum biofunctionalized with integrin binding mini-collagen FC3, cells
behave similar as on collagen I in terms of adhesion, spreading and migration. As recombinant minicollagen
FC3 is unhydroxylated, and thus highly specific for integrin ����, this integrin is fully
sufficient to induce this behaviour, while the participation of other collagen receptors is at most
ancillary under this conditions. 3
References
1. F. Rosenow, R. Ossig, D. Thormeyer, P. Gasmann, K. Schlüter, G. Brunner, J. Haier, J.A. Eble,
Neoplasia 2008, 10, 168-176.
2. J.A. Eble, S. Niland, T. Bracht, M. Mormann, J. Peter-Katalinic, G. Pohlentz, J. Stetefeld, FASEB J.
2009, 23, 2917-2927l.
3. S. Niland, C. Westerhausen, S.W. Schneider, M.F. Schneider, J.A. Eble, submitted to Biochem J.,
Bio-functionalization of a generic collagenous triple helix with the ���� integrin binding site allows
molecular force measurements.
33