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Title: Challenges and solutions for the analysis of in situ, in crystallo micro-spectrophotometric data

The particular challenge of the analysis of optical absorption and Raman spectroscopic data measured from protein crystals and how the SLS-APE software toolbox supports scientists in dealing with such data is described. Combining macromolecular crystallography with in crystallo micro-spectrophotometry yields valuable complementary information on the sample, including the redox states of metal cofactors, the identification of bound ligands and the onset and strength of undesired photochemistry, also known as radiation damage. However, the analysis and processing of the resulting data differs significantly from the approaches used for solution spectrophotometric data. The varying size and shape of the sample, together with the suboptimal sample environment, the lack of proper reference signals and the general influence of the X-ray beam on the sample have to be considered and carefully corrected for. In the present article, how to characterize and treat these sample-dependent artefacts in a reproducible manner is discussed and the SLS-APEin situ, in crystallo optical spectroscopy data-analysis toolbox is demonstrated.
Authors:
 [1] ;  [2] ;  [3] ;  [4] ;  [5]
  1. Paul Scherrer Institute, CH-5232 Villigen PSI (Switzerland)
  2. University of Essex, Wivenhoe Park, Colchester CO4 3SQ (United Kingdom)
  3. c/o DESY, Notkestrasse 85, D-22603 Hamburg (Germany)
  4. Brookhaven National Laboratory, Mail Stop 745, Upton, NY 11973 (United States)
  5. (Switzerland)
Publication Date:
OSTI Identifier:
22347735
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 1; Other Information: PMCID: PMC4304683; PMID: 25615857; PUBLISHER-ID: ba5221; OAI: oai:pubmedcentral.nih.gov:4304683; Copyright (c) Dworkowski et al. 2015; 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; BEAMS; CRYSTALS; DATA ANALYSIS; ENVIRONMENT; IRON; LIGANDS; MATHEMATICAL SOLUTIONS; PROTEINS; RAMAN SPECTROSCOPY; SHAPE; SIGNALS; SOLUTIONS; SPECTROPHOTOMETRY