Tracking X-ray-derived redox changes in crystals of a methylamine dehydrogenase/amicyanin complex using single-crystal UV/Vis microspectrophotometry.
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.
Pearson
Arwen R
AR
Department of Biochemistry, Molecular Biology and Biophysics, The University of Minnesota, Minneapolis, MN 55455, USA.
Pahl
Reinhard
R
Kovaleva
Elena G
EG
Davidson
Victor L
VL
Wilmot
Carrie M
CM
D001426:Bacterial Proteins / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D004307:Dose-Response Relationship, Radiation; D046912:Multiprotein Complexes / Q000737:chemistry / Q000528:radiation effects / Q000648:ultrastructure; D010084:Oxidation-Reduction / Q000528:radiation effects; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry* / Q000528:radiation effects* / Q000648:ultrastructure; D010231:Paracoccus denitrificans / Q000201:enzymology*; D011487:Protein Conformation / Q000528:radiation effects; D011829:Radiation Dosage; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D014965:X-Rays
D001426:Bacterial Proteins; D046912:Multiprotein Complexes; C017079:mauC protein, Methylobacterium extorquens; C021567:methylamine dehydrogenase; D000587:Oxidoreductases Acting on CH-NH Group Donors