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Biologische Systeme und Medizin Poster: Mi., 14:00–16:30 M-P199
Biological X-ray Absorption Spectroscopy in the 21st Century: Towards
automated Data-Collection and Analysis
Wolfram Meyer-Klaucke 1
1 EMBL Hamburg, Notkestr. 85, 22603 Hamburg, Germany
X-ray absorption spectroscopy allows the element specific characterization of metal
sites in biological systems, yielding information about the types and distances of coordinating
ligands as well as the oxidation state of the metal center. For multinuclear
proteins, the metalmetal distance can also be determined. In fact, the method can
be performed on samples in every state, including solutions and intact tissue. Energy
dependent measurements of the absorption coefficient require a tunable X-ray
source. Ideally, this is a synchrotron beamline equipped with a monochromator. The
tremendous improvements in this field allow for a high degree of automation during
the measurement. Increasingly, the data analysis becomes the bottle-neck. Recently,
the first packages for automated data-reduction became available[1]. On the way is the
automatic determination of metal binding motifs.
These options open the window towards high-throughput biological XAS. Some recent
ex-amples benefiting from these developments will be discussed in the second part of
the presen-tation, focusing on the metal specificity in the metallo-beta lactamase superfamily[2]
and pro-teins involved in metal regulation[3].
[1] Korbas, M., Fulla Marsa, D., Meyer-Klaucke, W. (2006) Kemp a program script
for automated biological XAS data reduction, Rev Sci Instr, accepted
Lippold, B, Meyer-Klaucke, W, Meyer, T, and Henkel, G (2005) Towards an automated
quality control of XAS data. J Synchrotron Radiat 12, 45-52.
[2] Schilling, O, Vogel, A, Kostelecky, B, Natal da Luz, H, Spemann, D, Spath, B,
March-felder, A, Troger, W, and Meyer-Klaucke, W (2005) Zinc- and iron-dependent
cytosolic metallo-beta-lactamase domain proteins exhibit similar zinc-binding affinities,
independ-ent of an atypical glutamate at the metal-binding site. Biochem J 385, 145-
153.
Schilling, O, Wenzel, N, Naylor, M, Vogel, A, Crowder, M, Makaroff, C, and Meyer-
Klaucke, W, Flexible metal binding of the metallo-beta-lactamase domain: glyoxalase
II incorporates iron, manganese, and zinc in vivo. (2003) Biochemistry 42, 11777-11786
[3] Pohl, E, Haller, JC, Mijovilovich, A, Meyer-Klaucke, W, Garman, E, and Vasil,
ML, (2003) Architecture of a protein central to iron homeostasis: crystal structure and
spectroscopic analysis of the ferric uptake regulator. Mol Microbiol 47, 903-915.