Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography.

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Abstract

Authors: Uwe Weierstall,Daniel James, Chong Wang, Thomas A. White, Dingjie Wang, Wei Liu, John C. H. Spence, R. Bruce Doak, Garrett Nelson, Petra Fromme, Raimund Fromme, Ingo Grotjohann, Christopher Kupitz, Nadia A. Zatsepin, Haiguang Liu, Shibom Basu, Daniel Wacker, Gye Won Han, Vsevolod Katritch, Sébastien Boutet, Marc Messerschmidt, Garth J. Williams, Jason E. Koglin, M. Marvin Seibert, Markus Klinker,Cornelius Gati, Robert L. Shoeman, Anton Barty, Henry N. Chapman, Richard A. Kirian, Kenneth R. Beyerlein, Raymond C. Stevens, Dianfan Li, Syed T. A. Shah, Nicole Howe, Martin Caffrey & Vadim Cherezov

Lipidic cubic phase (LCP) crystallization has proven successful for high-resolution structure determination of challenging membrane proteins. Here we present a technique for extruding gel-like LCP with embedded membrane protein microcrystals, providing a continuously renewed source of material for serial femtosecond crystallography. Data collected from sub-10-μm-sized crystals produced with less than 0.5 mg of purified protein yield structural insights regarding cyclopamine binding to the Smoothened receptor.

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Joey Olmos (Rice)
April 15th, 2015