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

Discipline Tags:



Technical Tags: