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Title: Insights into the mechanism of X-ray-induced disulfide-bond cleavage in lysozyme crystals based on EPR, optical absorption and X-ray diffraction studies

Electron paramagnetic resonance (EPR) and online UV–visible absorption microspectrophotometry with X-ray crystallography have been used in a complementary manner to follow X-ray-induced disulfide-bond cleavage, to confirm a multi-track radiation-damage process and to develop a model of that process. Electron paramagnetic resonance (EPR) and online UV–visible absorption microspectrophotometry with X-ray crystallography have been used in a complementary manner to follow X-ray-induced disulfide-bond cleavage. Online UV–visible spectroscopy showed that upon X-irradiation, disulfide radicalization appeared to saturate at an absorbed dose of approximately 0.5–0.8 MGy, in contrast to the saturating dose of ∼0.2 MGy observed using EPR at much lower dose rates. The observations suggest that a multi-track model involving product formation owing to the interaction of two separate tracks is a valid model for radiation damage in protein crystals. The saturation levels are remarkably consistent given the widely different experimental parameters and the range of total absorbed doses studied. The results indicate that even at the lowest doses used for structural investigations disulfide bonds are already radicalized. Multi-track considerations offer the first step in a comprehensive model of radiation damage that could potentially lead to a combined computational and experimental approach to identifying when damage is likely to be present, to quantitatemore » it and to provide the ability to recover the native unperturbed structure.« less
Authors:
 [1] ; ;  [2] ;  [3] ;  [4] ;  [1] ;  [5] ;  [2] ;  [5]
  1. Hauptman–Woodward Medical Research Institute, 700 Ellicott Street, Buffalo, NY 14086 (United States)
  2. University of Rochester Medical Center, Rochester, NY 14642 (United States)
  3. University of Oxford, South Parks Road, Oxford, Oxfordshire OX1 3QU (United Kingdom)
  4. Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom)
  5. (United States)
Publication Date:
OSTI Identifier:
22347802
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 69; Journal Issue: Pt 12; Other Information: PMCID: PMC3852651; PMID: 24311579; PUBLISHER-ID: kw5071; OAI: oai:pubmedcentral.nih.gov:3852651; Copyright (c) Sutton, Black et al. 2013; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; ABSORPTION; CRYSTALLOGRAPHY; CRYSTALS; DAMAGE; ELECTRON SPIN RESONANCE; ELECTRONS; INTERACTIONS; IRRADIATION; LYSOZYME; PARTICLE TRACKS; PROTEINS; RADIATION EFFECTS; SPECTROSCOPY; X-RAY DIFFRACTION