Journal of Synchrotron Radiation

Volume 14, Part 1 (January 2007)

radiation damage

J. Synchrotron Rad. (2007). 14, 92-98 [ doi:10.1107/S0909049506051259 ]

Tracking X-ray-derived redox changes in crystals of a methylamine dehydrogenase/amicyanin complex using single-crystal UV/Vis microspectrophotometry

A. R. Pearson, R. Pahl, E. G. Kovaleva, V. L. Davidson and C. M. Wilmot

Abstract: X-ray exposure during crystallographic data collection can result in unintended redox changes in proteins containing functionally important redox centers. In order to directly monitor X-ray-derived redox changes in trapped oxidative half-reaction intermediates of Paracoccus denitrificans methylamine dehydrogenase, a commercially available single-crystal UV/Vis microspectrophotometer was installed on-line at the BioCARS beamline 14-BM-C at the Advanced Photon Source, Argonne, USA. Monitoring the redox state of the intermediates during X-ray exposure permitted the creation of a general multi-crystal data collection strategy to generate true structures of each redox intermediate.

Keywords: single-crystal microspectrophotometry; reaction intermediates; structural enzymology; tryptophan tryptophylquinone; methylamine dehydrogenase.


	GIF image (476.2 kbytes) [ doi:10.1107/S0909049506051259/ms5001sup1.gif ] Movie showing a series of single-crystal spectra recorded as a TTQ_OX / Cu^2+ MADH/amicyanin crystal is rotated about angle phi. The anisotropic nature of single-crystal spectra is clearly observed.

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