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Structure determination of an integral membrane protein at room temperature from crystals in situ
DOI:
10.1107/S139900471500423X
PMID:
26057664
Authors:
Danny
Axford
(Diamond Light Source)
,
James
Foadi
(Diamond Light Source)
,
Nien-jen
Hu
(Diamond Light Source)
,
Hassan
Choudhury
(Diamond Light Source)
,
So
Iwata
(Diamond Light Source)
,
Konstantinos
Beis
(Diamond Light Source)
,
Gwyndaf
Evans
(Diamond Light Source)
,
Yilmaz
Alguel
(Imperial College MSF, Diamond Light Source)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Acta Crystallographica Section D Biological Crystallography
, VOL 71
State:
Published (Approved)
Published:
June 2015

Abstract: 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 A˚ 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.
Journal Keywords: Crystallization; Crystals; Diffusion; Membrane Proteins; Plates; Potentials; Resolution; Synchrotrons; X-Ray Diffraction
Subject Areas:
Technique Development
Diamond Offline Facilities:
Membrane Protein Laboratory (MPL)
Instruments:
I24-Microfocus Macromolecular Crystallography
Added On:
23/05/2015 16:32
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