Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography

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

doi:10.1038/ncomms4309

Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography

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

Research output: Contribution to journal › Article › peer-review

487 Scopus citations

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 language	English
Article number	3309
Journal	Nature Communications
Volume	5
DOIs	https://doi.org/10.1038/ncomms4309
State	Published - 14 Feb 2014
Externally published	Yes

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10.1038/ncomms4309

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Weierstall, U., James, D., Wang, C., White, T. A., Wang, D., Liu, W., Spence, J. C. H., Bruce Doak, R., Nelson, G., Fromme, P., Fromme, R., Grotjohann, I., Kupitz, C., Zatsepin, N. A., Liu, H., Basu, S., Wacker, D., Won Han, G., Katritch, V., ... Cherezov, V. (2014). Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography. Nature Communications, 5, Article 3309. https://doi.org/10.1038/ncomms4309

@article{05ef6db5a3214b44ae0f58ab2c8b3b94,

title = "Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography",

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.",

author = "Uwe Weierstall and Daniel James and Chong Wang and White, \{Thomas A.\} and Dingjie Wang and Wei Liu and Spence, \{John C.H.\} and \{Bruce Doak\}, R. and Garrett Nelson and Petra Fromme and Raimund Fromme and Ingo Grotjohann and Christopher Kupitz and Zatsepin, \{Nadia A.\} and Haiguang Liu and Shibom Basu and Daniel Wacker and \{Won Han\}, Gye and Vsevolod Katritch and S{\'e}bastien Boutet and Marc Messerschmidt and Williams, \{Garth J.\} and Koglin, \{Jason E.\} and \{Marvin Seibert\}, M. and Markus Klinker and Cornelius Gati and Shoeman, \{Robert L.\} and Anton Barty and Chapman, \{Henry N.\} and Kirian, \{Richard A.\} and Beyerlein, \{Kenneth R.\} and Stevens, \{Raymond C.\} and Dianfan Li and Shah, \{Syed T.A.\} and Nicole Howe and Martin Caffrey and Vadim Cherezov",

note = "Funding Information: Experiments were carried out at the Linac Coherent Light Source, a national user facility operated by Stanford University on behalf of the U.S. Department of Energy (DOE), Office of Basic Energy Sciences. The work was supported in part by the National Institutes of Health Common Fund in Structural Biology grants P50 GM073197 (V.C. and R.C.S.), P50 GM073210 (M.C.), R01 GM095583 (P.F.) and NIGMS PSI:Biology grants U54 GM094618 (V.C. and R.C.S.), U54 GM094599 (P.F.), U54 GM094625 (M.C.). We acknowledge support from the National Science Foundation (awards MCB 1021557 and MCB 1120997) (J.C.H.S., U.W., P.F., R.B.D.) and its BioXFEL Science and Technology Center (NSF 1231306) and the Science Foundation Ireland (12/IA/1255). We further acknowledge support from the Helmholtz Gesellschaft and the Max Planck Gesellschaft (R.L.S.). Special thanks to T. Trinh and M. Chu for the help with baculovirus expression, F. Siu for help with sample preparation, and A. Walker for assistance with manuscript preparation. We are grateful to Richard Flubacher and the machinists of the ASU Mechanical Instrument Shop for their skill in fabricating injector components.",

year = "2014",

month = feb,

day = "14",

doi = "10.1038/ncomms4309",

language = "English",

volume = "5",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Research",

}

Weierstall, U, James, D, Wang, C, White, TA, Wang, D, Liu, W, Spence, JCH, Bruce Doak, R, Nelson, G, Fromme, P, Fromme, R, Grotjohann, I, Kupitz, C, Zatsepin, NA, 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, RA, 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

TY - JOUR

T1 - Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography

AU - Weierstall, Uwe

AU - James, Daniel

AU - Wang, Chong

AU - White, Thomas A.

AU - Wang, Dingjie

AU - Liu, Wei

AU - Spence, John C.H.

AU - Bruce Doak, R.

AU - Nelson, Garrett

AU - Fromme, Petra

AU - Fromme, Raimund

AU - Grotjohann, Ingo

AU - Kupitz, Christopher

AU - Zatsepin, Nadia A.

AU - Liu, Haiguang

AU - Basu, Shibom

AU - Wacker, Daniel

AU - Won Han, Gye

AU - Katritch, Vsevolod

AU - Boutet, Sébastien

AU - Messerschmidt, Marc

AU - Williams, Garth J.

AU - Koglin, Jason E.

AU - Marvin Seibert, M.

AU - Klinker, Markus

AU - Gati, Cornelius

AU - Shoeman, Robert L.

AU - Barty, Anton

AU - Chapman, Henry N.

AU - Kirian, Richard A.

AU - Beyerlein, Kenneth R.

AU - Stevens, Raymond C.

AU - Li, Dianfan

AU - Shah, Syed T.A.

AU - Howe, Nicole

AU - Caffrey, Martin

AU - Cherezov, Vadim

N1 - Funding Information: Experiments were carried out at the Linac Coherent Light Source, a national user facility operated by Stanford University on behalf of the U.S. Department of Energy (DOE), Office of Basic Energy Sciences. The work was supported in part by the National Institutes of Health Common Fund in Structural Biology grants P50 GM073197 (V.C. and R.C.S.), P50 GM073210 (M.C.), R01 GM095583 (P.F.) and NIGMS PSI:Biology grants U54 GM094618 (V.C. and R.C.S.), U54 GM094599 (P.F.), U54 GM094625 (M.C.). We acknowledge support from the National Science Foundation (awards MCB 1021557 and MCB 1120997) (J.C.H.S., U.W., P.F., R.B.D.) and its BioXFEL Science and Technology Center (NSF 1231306) and the Science Foundation Ireland (12/IA/1255). We further acknowledge support from the Helmholtz Gesellschaft and the Max Planck Gesellschaft (R.L.S.). Special thanks to T. Trinh and M. Chu for the help with baculovirus expression, F. Siu for help with sample preparation, and A. Walker for assistance with manuscript preparation. We are grateful to Richard Flubacher and the machinists of the ASU Mechanical Instrument Shop for their skill in fabricating injector components.

PY - 2014/2/14

Y1 - 2014/2/14

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/84894037590

U2 - 10.1038/ncomms4309

DO - 10.1038/ncomms4309

M3 - Article

C2 - 24525480

AN - SCOPUS:84894037590

SN - 2041-1723

VL - 5

JO - Nature Communications

JF - Nature Communications

M1 - 3309

ER -

Lipidic cubic phase injector facilitates membrane protein serial femtosecond crystallography

Abstract

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