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Title: Structure determination of an integral membrane protein at room temperature from crystals in situ

The X-ray structure determination of an integral membrane protein using synchrotron diffraction data measured in situ at room temperature is demonstrated. The structure determination of an integral membrane protein using synchrotron X-ray diffraction data collected at room temperature directly in vapour-diffusion crystallization plates (in situ) is demonstrated. Exposing the crystals in situ eliminates manual sample handling and, since it is performed at room temperature, removes the complication of cryoprotection and potential structural anomalies induced by sample cryocooling. Essential to the method is the ability to limit radiation damage by recording a small amount of data per sample from many samples and subsequently assembling the resulting data sets using specialized software. The validity of this procedure is established by the structure determination of Haemophilus influenza TehA at 2.3 Å resolution. The method presented offers an effective protocol for the fast and efficient determination of membrane-protein structures at room temperature using third-generation synchrotron beamlines.
Authors:
 [1] ;  [1] ;  [2] ; ;  [1] ;  [2] ;  [2] ;  [1] ;  [2] ;  [2] ;  [2] ;  [3] ;  [1] ;  [2] ;  [2] ;  [1] ;  [1] ;  [2] ;  [2]
  1. Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom)
  2. (United Kingdom)
  3. (Japan)
Publication Date:
OSTI Identifier:
22389067
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section D: Biological Crystallography; Journal Volume: 71; Journal Issue: Pt 6; Other Information: PMCID: PMC4461203; PMID: 26057664; PUBLISHER-ID: tz5076; OAI: oai:pubmedcentral.nih.gov:4461203; Copyright (c) Axford 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; CRYSTALLIZATION; CRYSTALS; DIFFUSION; MEMBRANE PROTEINS; PLATES; POTENTIALS; RESOLUTION; SYNCHROTRONS; X-RAY DIFFRACTION