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 A. 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 C.H. 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 A. 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 C.H. Spence, Raimund Fromme, Uwe Weierstall, Petra Fromme

Arizona State University

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

85 Citations (Scopus)

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
Pages (from-to)	421-430
Number of pages	10
Journal	IUCrJ
Volume	2
DOIs	https://doi.org/10.1107/S2052252515009811
Publication status	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

Chemistry(all)
Biochemistry
Materials Science(all)
Condensed Matter Physics

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Conrad, C. E., Basu, S., James, D., Wang, D., Schaffer, A., Roy-Chowdhury, S., Zatsepin, N. A., 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

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

In: IUCrJ, Vol. 2, 01.07.2015, p. 421-430.

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

Conrad, CE, Basu, S, James, D, Wang, D, Schaffer, A, Roy-Chowdhury, S, Zatsepin, NA, 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, JCH, 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

Conrad, Chelsie E. ; Basu, Shibom ; James, Daniel ; Wang, Dingjie ; Schaffer, Alexander ; Roy-Chowdhury, Shatabdi ; Zatsepin, Nadia A. ; Aquila, Andrew ; Coe, Jesse ; Gati, Cornelius ; Hunter, Mark S. ; Koglin, Jason E. ; Kupitz, Christopher ; Nelson, Garrett ; Subramanian, Ganesh ; White, Thomas A. ; Zhao, Yun ; Zook, James ; Boutet, Sébastien ; Cherezov, Vadim ; Spence, John C.H. ; Fromme, Raimund ; Weierstall, Uwe ; Fromme, Petra. / A novel inert crystal delivery medium for serial femtosecond crystallography. In: IUCrJ. 2015 ; Vol. 2. pp. 421-430.

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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 C.H.

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AU - Weierstall, Uwe

AU - Fromme, Petra

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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.

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