Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography

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

Research output: Contribution to journalArticle

296 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article number3309
JournalNature Communications
Volume5
DOIs
StatePublished - Feb 14 2014

Fingerprint

Crystallography
injectors
crystallography
Membrane Proteins
membranes
proteins
Microcrystals
Crystallization
extruding
Gels
microcrystals
Crystals
gels
crystallization
Proteins
high resolution
crystals
cyclopamine
Smoothened Receptor

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

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

In: Nature Communications, Vol. 5, 3309, 14.02.2014.

Research output: Contribution to journalArticle

Weierstall, U, James, D, Wang, C, White, TA, Wang, D, Liu, W, Spence, J, Bruce Doak, R, Nelson, G, Fromme, P, Fromme, R, Grotjohann, I, Kupitz, C, Zatsepin, N, Liu, H, Basu, S, Wacker, D, Won Han, G, Katritch, V, Boutet, S, Messerschmidt, M, Williams, GJ, Koglin, JE, Marvin Seibert, M, Klinker, M, Gati, C, Shoeman, RL, Barty, A, Chapman, HN, Kirian, R, Beyerlein, KR, Stevens, RC, Li, D, Shah, STA, Howe, N, Caffrey, M & Cherezov, V 2014, 'Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography', Nature Communications, vol. 5, 3309. https://doi.org/10.1038/ncomms4309
Weierstall, Uwe ; James, Daniel ; Wang, Chong ; White, Thomas A. ; Wang, Dingjie ; Liu, Wei ; Spence, John ; Bruce Doak, R. ; Nelson, Garrett ; Fromme, Petra ; Fromme, Raimund ; Grotjohann, Ingo ; Kupitz, Christopher ; Zatsepin, Nadia ; Liu, Haiguang ; Basu, Shibom ; Wacker, Daniel ; Won Han, Gye ; Katritch, Vsevolod ; Boutet, Sébastien ; Messerschmidt, Marc ; Williams, Garth J. ; Koglin, Jason E. ; Marvin Seibert, M. ; Klinker, Markus ; Gati, Cornelius ; Shoeman, Robert L. ; Barty, Anton ; Chapman, Henry N. ; Kirian, Richard ; Beyerlein, Kenneth R. ; Stevens, Raymond C. ; Li, Dianfan ; Shah, Syed T A ; Howe, Nicole ; Caffrey, Martin ; Cherezov, Vadim. / Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography. In: Nature Communications. 2014 ; Vol. 5.
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