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Monitoring and validating active site redox states in protein crystals

DOI: 10.1016/j.bbapap.2010.12.017 DOI Help
PMID: 21215826 PMID Help

Authors: Svetlana Antonyuk (University of Liverpool) , Mike Hough (University of Essex)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Biochimica Et Biophysica Acta (bba) - Proteins And Proteomics

State: Published (Approved)
Published: January 2011
Diamond Proposal Number(s): 1215

Abstract: High resolution protein crystallography using synchrotron radiation is one of the most powerful tools in modern biology. Improvements in resolution have arisen from the use of X-ray beamlines with higher brightness and flux and the development of advanced detectors. However, it is increasingly recognised that the benefits brought by these advances have an associated cost, namely deleterious effects of X-ray radiation on the sample (radiation damage). In particular, X-ray induced reduction and damage to redox centres has been shown to occur much more rapidly than other radiation damage effects, such as loss of resolution or damage to disulphide bridges. Selection of an appropriate combination of in-situ single crystal spectroscopies during crystallographic experiments, such as UV-visible absorption and X-ray absorption spectroscopy (XAFS), allows for effective monitoring of redox states in protein crystals in parallel with structure determination. Such approaches are also essential in cases where catalytic intermediate species are generated by exposure to the X-ray beam. In this article, we provide a number of examples in which multiple single crystal spectroscopies have been key to understanding the redox status of Fe and Cu centres in crystal structures. This article is part of a Special Issue entitled: Protein Structure and Function in the Crystalline State.

Journal Keywords: Catalytic; Copper; Crystallography; X-Ray; Cytochrome; Ferritins; Iron; Metalloproteins; Models; Molecular; Nitrite; Oxidation-Reduction; Reproducibility; Spectrophotometry; Ultraviolet; Spectrum; X-Ray Absorption Spectroscopy

Subject Areas: Biology and Bio-materials

Instruments: I03-Macromolecular Crystallography