Open AccessStructure determination of an integral membrane protein at room temperature from crystals in situ
Author(s) -
Axford Danny,
Foadi James,
Hu NienJen,
Choudhury Hassanul Ghani,
Iwata So,
Beis Konstantinos,
Evans Gwyndaf,
Alguel Yilmaz
Publication year - 2015
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s139900471500423x
Subject(s) - synchrotron radiation , synchrotron , in situ , crystallization , materials science , resolution (logic) , diffraction , diffusion , sample (material) , membrane , analytical chemistry (journal) , optics , x ray crystallography , crystallography , chemistry , chromatography , physics , computer science , thermodynamics , biochemistry , organic chemistry , artificial intelligence
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.