A novel inert crystal delivery medium for serial femtosecond crystallography

Chelsie E. Conrad; Shibom Basu; Daniel James; Dingjie Wang; Alexander Schaffer; Shatabdi Roy-Chowdhury; Nadia Zatsepin; Andrew Aquila; Jesse Coe; Cornelius Gati; Mark S. Hunter; Jason E. Koglin; Christopher Kupitz; Garrett Nelson; Ganesh Subramanian; Thomas A. White; Yun Zhao; James Zook; Sébastien Boutet; Vadim Cherezov; John Spence; Raimund Fromme; Uwe Weierstall; Petra Fromme

doi:10.1107/S2052252515009811

A novel inert crystal delivery medium for serial femtosecond crystallography

Chelsie E. Conrad, Shibom Basu, Daniel James, Dingjie Wang, Alexander Schaffer, Shatabdi Roy-Chowdhury, Nadia Zatsepin, Andrew Aquila, Jesse Coe, Cornelius Gati, Mark S. Hunter, Jason E. Koglin, Christopher Kupitz, Garrett Nelson, Ganesh Subramanian, Thomas A. White, Yun Zhao, James Zook, Sébastien Boutet, Vadim CherezovJohn Spence, Raimund Fromme, Uwe Weierstall, Petra Fromme

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

113 Scopus citations

Abstract

Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 μg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.

Original language	English (US)
Pages (from-to)	421-430
Number of pages	10
Journal	IUCrJ
Volume	2
DOIs	https://doi.org/10.1107/S2052252515009811
State	Published - Jul 1 2015

Keywords

coherent X-ray diffractive imaging
femtosecond studies
free-electron laser
membrane proteins
nanocrystals
protein complexes
serial femtosecond crystallography
viscous crystal delivery

ASJC Scopus subject areas

General Chemistry
Biochemistry
General Materials Science
Condensed Matter Physics

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10.1107/S2052252515009811

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Conrad, C. E., Basu, S., James, D., Wang, D., Schaffer, A., Roy-Chowdhury, S., Zatsepin, N., Aquila, A., Coe, J., Gati, C., Hunter, M. S., Koglin, J. E., Kupitz, C., Nelson, G., Subramanian, G., White, T. A., Zhao, Y., Zook, J., Boutet, S., ... Fromme, P. (2015). A novel inert crystal delivery medium for serial femtosecond crystallography. IUCrJ, 2, 421-430. https://doi.org/10.1107/S2052252515009811

Conrad, CE, Basu, S, James, D, Wang, D, Schaffer, A, Roy-Chowdhury, S, Zatsepin, N, Aquila, A, Coe, J, Gati, C, Hunter, MS, Koglin, JE, Kupitz, C, Nelson, G, Subramanian, G, White, TA, Zhao, Y, Zook, J, Boutet, S, Cherezov, V, Spence, J, Fromme, R , Weierstall, U & Fromme, P 2015, 'A novel inert crystal delivery medium for serial femtosecond crystallography', IUCrJ, vol. 2, pp. 421-430. https://doi.org/10.1107/S2052252515009811

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abstract = "Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 {\AA} resolution using 300 μg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.",

keywords = "coherent X-ray diffractive imaging, femtosecond studies, free-electron laser, membrane proteins, nanocrystals, protein complexes, serial femtosecond crystallography, viscous crystal delivery",

author = "Conrad, {Chelsie E.} and Shibom Basu and Daniel James and Dingjie Wang and Alexander Schaffer and Shatabdi Roy-Chowdhury and Nadia Zatsepin and Andrew Aquila and Jesse Coe and Cornelius Gati and Hunter, {Mark S.} and Koglin, {Jason E.} and Christopher Kupitz and Garrett Nelson and Ganesh Subramanian and White, {Thomas A.} and Yun Zhao and James Zook and S{\'e}bastien Boutet and Vadim Cherezov and John Spence and Raimund Fromme and Uwe Weierstall and Petra Fromme",

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doi = "10.1107/S2052252515009811",

language = "English (US)",

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AU - Conrad, Chelsie E.

AU - Basu, Shibom

AU - James, Daniel

AU - Wang, Dingjie

AU - Schaffer, Alexander

AU - Roy-Chowdhury, Shatabdi

AU - Zatsepin, Nadia

AU - Aquila, Andrew

AU - Coe, Jesse

AU - Gati, Cornelius

AU - Hunter, Mark S.

AU - Koglin, Jason E.

AU - Kupitz, Christopher

AU - Nelson, Garrett

AU - Subramanian, Ganesh

AU - White, Thomas A.

AU - Zhao, Yun

AU - Zook, James

AU - Boutet, Sébastien

AU - Cherezov, Vadim

AU - Spence, John

AU - Fromme, Raimund

AU - Weierstall, Uwe

AU - Fromme, Petra

PY - 2015/7/1

Y1 - 2015/7/1

N2 - Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 μg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.

AB - Serial femtosecond crystallography (SFX) has opened a new era in crystallo-graphy by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 μg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.

KW - coherent X-ray diffractive imaging

KW - femtosecond studies

KW - free-electron laser

KW - membrane proteins

KW - nanocrystals

KW - protein complexes

KW - serial femtosecond crystallography

KW - viscous crystal delivery

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A novel inert crystal delivery medium for serial femtosecond crystallography

Abstract

Keywords

ASJC Scopus subject areas

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Phycocyanin structure from T. elongatus at 2.5-A from XFEL using a viscous delivery medium for serial femtosecond crystallography

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A novel inert crystal delivery medium for serial femtosecond crystallography

Abstract

Keywords

ASJC Scopus subject areas

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Phycocyanin structure from T. elongatus at 2.5-A from XFEL using a viscous delivery medium for serial femtosecond crystallography

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