High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source

Jose M. Martin-Garcia; Lan Zhu; Derek Mendez; Ming Yue Lee; Eugene Chun; Chufeng Li; Hao Hu; Ganesh Subramanian; David Kissick; Craig Ogata; Robert Henning; Andrii Ishchenko; Zachary Dobson; Shangji Zhang; Uwe Weierstall; John C.H. Spence; Petra Fromme; Nadia A. Zatsepin; Robert F. Fischetti; Vadim Cherezov; Wei Liu

doi:10.1107/S205225251900263X

High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source

Jose M. Martin-Garcia, Lan Zhu, Derek Mendez, Ming Yue Lee, Eugene Chun, Chufeng Li, Hao Hu, Ganesh Subramanian, David Kissick, Craig Ogata, Robert Henning, Andrii Ishchenko, Zachary Dobson, Shangji Zhang, Uwe Weierstall, John C.H. Spence, Petra Fromme, Nadia A. Zatsepin, Robert F. Fischetti, Vadim CherezovWei Liu

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

35 Scopus citations

Abstract

Since the first successful serial crystallography (SX) experiment at a synchrotron radiation source, the popularity of this approach has continued to grow showing that third-generation synchrotrons can be viable alternatives to scarce X-ray free-electron laser sources. Synchrotron radiation flux may be increased ∼100 times by a moderate increase in the bandwidth ('pink beam' conditions) at some cost to data analysis complexity. Here, we report the first high-viscosity injector-based pink-beam SX experiments. The structures of proteinase K (PK) and A 2A adenosine receptor (A 2A AR) were determined to resolutions of 1.8 and 4.2Å using 4 and 24 consecutive 100ps X-ray pulse exposures, respectively. Strong PK data were processed using existing Laue approaches, while weaker A 2A AR data required an alternative data-processing strategy. This demonstration of the feasibility presents new opportunities for time-resolved experiments with microcrystals to study structural changes in real time at pink-beam synchrotron beamlines worldwide.

Original language	English (US)
Pages (from-to)	412-425
Number of pages	14
Journal	IUCrJ
Volume	6
DOIs	https://doi.org/10.1107/S205225251900263X
State	Published - 2019

Keywords

X-ray crystallography
injector-based serial crystallography
membrane proteins
pink-beam serial crystallography
protein structures
structural biology
structure determination
third-generation synchrotrons

ASJC Scopus subject areas

General Chemistry
Biochemistry
General Materials Science
Condensed Matter Physics

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10.1107/S205225251900263X

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Martin-Garcia, J. M., Zhu, L., Mendez, D., Lee, M. Y., Chun, E., Li, C., Hu, H., Subramanian, G., Kissick, D., Ogata, C., Henning, R., Ishchenko, A., Dobson, Z., Zhang, S., Weierstall, U., Spence, J. C. H., Fromme, P., Zatsepin, N. A., Fischetti, R. F., ... Liu, W. (2019). High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source. IUCrJ, 6, 412-425. https://doi.org/10.1107/S205225251900263X

Martin-Garcia, JM, Zhu, L, Mendez, D, Lee, MY, Chun, E, Li, C, Hu, H, Subramanian, G, Kissick, D, Ogata, C, Henning, R, Ishchenko, A, Dobson, Z, Zhang, S, Weierstall, U, Spence, JCH, Fromme, P, Zatsepin, NA, Fischetti, RF, Cherezov, V & Liu, W 2019, 'High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source', IUCrJ, vol. 6, pp. 412-425. https://doi.org/10.1107/S205225251900263X

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title = "High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source",

abstract = "Since the first successful serial crystallography (SX) experiment at a synchrotron radiation source, the popularity of this approach has continued to grow showing that third-generation synchrotrons can be viable alternatives to scarce X-ray free-electron laser sources. Synchrotron radiation flux may be increased ∼100 times by a moderate increase in the bandwidth ('pink beam' conditions) at some cost to data analysis complexity. Here, we report the first high-viscosity injector-based pink-beam SX experiments. The structures of proteinase K (PK) and A 2A adenosine receptor (A 2A AR) were determined to resolutions of 1.8 and 4.2{\AA} using 4 and 24 consecutive 100ps X-ray pulse exposures, respectively. Strong PK data were processed using existing Laue approaches, while weaker A 2A AR data required an alternative data-processing strategy. This demonstration of the feasibility presents new opportunities for time-resolved experiments with microcrystals to study structural changes in real time at pink-beam synchrotron beamlines worldwide.",

keywords = "X-ray crystallography, injector-based serial crystallography, membrane proteins, pink-beam serial crystallography, protein structures, structural biology, structure determination, third-generation synchrotrons",

author = "Martin-Garcia, {Jose M.} and Lan Zhu and Derek Mendez and Lee, {Ming Yue} and Eugene Chun and Chufeng Li and Hao Hu and Ganesh Subramanian and David Kissick and Craig Ogata and Robert Henning and Andrii Ishchenko and Zachary Dobson and Shangji Zhang and Uwe Weierstall and Spence, {John C.H.} and Petra Fromme and Zatsepin, {Nadia A.} and Fischetti, {Robert F.} and Vadim Cherezov and Wei Liu",

note = "Funding Information: This work was supported by the Centre for Applied Structural Discovery at the Biodesign Institute at Arizona State University (W. Liu, P. Fromme, U. Weierstall, J. C. H. Spence, J. M. Martin-Garcia, N. A. Zatsepin, L. Zhu, M.-Y. Lee, E. Chun, G. Subramanian), a Mayo Clinic ASU Collaborative Seed Grant Award (W. Liu), the Flinn Foundation Seed Grant (W. Liu) and No. 1991 (P. Fromme, J. C. H. Spence, and J. M. Martin-Garcia), the STC Program of the National Science Foundation through BioXFEL (No. 1231306; W. Liu, P. Fromme, U. Weierstall, J. C. H. Spence, N. A. Zatsepin, D. Mendez, C. Li, G. Subramanian), National Science Foundation BIO ABI grant No. 1565180 (N. A. Zatsepin, D. Mendez, U. Weierstall, and J. C. H. Spence), and the National Institutes of Health (NIH) grants R21DA042298 (W. Liu), R01GM124152 (W. Liu), R01GM095583 (P. Fromme) and R35GM127086 (V. Cherezov and A. Ishchenko). Use of BioCARS was supported by the National Institute of General Medical Sciences of the National Institutes of Health under grant number R24GM111072. Time-resolved setup at sector 14 was funded in part through a collaboration with Philip Anfinrud (NIH/ NIDDK). Use of the GM/CA at the Advanced Photon Source, Argonne National Laboratory was supported by the National Cancer Institute (ACB-12002; R. F. Fischetti, D. Kissick, and C. Ogata) and the National Institute of General Medical Sciences (AGM12006; R. F. Fischetti, D. Kissick, and C. Ogata). Use of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory, was supported under Contract No. DE-AC02-06CH11357. Publisher Copyright: {\textcopyright} Martin-Garcia et al. 2019.",

year = "2019",

doi = "10.1107/S205225251900263X",

language = "English (US)",

volume = "6",

pages = "412--425",

journal = "IUCrJ",

issn = "2052-2525",

publisher = "International Union of Crystallography",

}

TY - JOUR

T1 - High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source

AU - Martin-Garcia, Jose M.

AU - Zhu, Lan

AU - Mendez, Derek

AU - Lee, Ming Yue

AU - Chun, Eugene

AU - Li, Chufeng

AU - Hu, Hao

AU - Subramanian, Ganesh

AU - Kissick, David

AU - Ogata, Craig

AU - Henning, Robert

AU - Ishchenko, Andrii

AU - Dobson, Zachary

AU - Zhang, Shangji

AU - Weierstall, Uwe

AU - Spence, John C.H.

AU - Fromme, Petra

AU - Zatsepin, Nadia A.

AU - Fischetti, Robert F.

AU - Cherezov, Vadim

AU - Liu, Wei

N1 - Funding Information: This work was supported by the Centre for Applied Structural Discovery at the Biodesign Institute at Arizona State University (W. Liu, P. Fromme, U. Weierstall, J. C. H. Spence, J. M. Martin-Garcia, N. A. Zatsepin, L. Zhu, M.-Y. Lee, E. Chun, G. Subramanian), a Mayo Clinic ASU Collaborative Seed Grant Award (W. Liu), the Flinn Foundation Seed Grant (W. Liu) and No. 1991 (P. Fromme, J. C. H. Spence, and J. M. Martin-Garcia), the STC Program of the National Science Foundation through BioXFEL (No. 1231306; W. Liu, P. Fromme, U. Weierstall, J. C. H. Spence, N. A. Zatsepin, D. Mendez, C. Li, G. Subramanian), National Science Foundation BIO ABI grant No. 1565180 (N. A. Zatsepin, D. Mendez, U. Weierstall, and J. C. H. Spence), and the National Institutes of Health (NIH) grants R21DA042298 (W. Liu), R01GM124152 (W. Liu), R01GM095583 (P. Fromme) and R35GM127086 (V. Cherezov and A. Ishchenko). Use of BioCARS was supported by the National Institute of General Medical Sciences of the National Institutes of Health under grant number R24GM111072. Time-resolved setup at sector 14 was funded in part through a collaboration with Philip Anfinrud (NIH/ NIDDK). Use of the GM/CA at the Advanced Photon Source, Argonne National Laboratory was supported by the National Cancer Institute (ACB-12002; R. F. Fischetti, D. Kissick, and C. Ogata) and the National Institute of General Medical Sciences (AGM12006; R. F. Fischetti, D. Kissick, and C. Ogata). Use of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory, was supported under Contract No. DE-AC02-06CH11357. Publisher Copyright: © Martin-Garcia et al. 2019.

PY - 2019

Y1 - 2019

N2 - Since the first successful serial crystallography (SX) experiment at a synchrotron radiation source, the popularity of this approach has continued to grow showing that third-generation synchrotrons can be viable alternatives to scarce X-ray free-electron laser sources. Synchrotron radiation flux may be increased ∼100 times by a moderate increase in the bandwidth ('pink beam' conditions) at some cost to data analysis complexity. Here, we report the first high-viscosity injector-based pink-beam SX experiments. The structures of proteinase K (PK) and A 2A adenosine receptor (A 2A AR) were determined to resolutions of 1.8 and 4.2Å using 4 and 24 consecutive 100ps X-ray pulse exposures, respectively. Strong PK data were processed using existing Laue approaches, while weaker A 2A AR data required an alternative data-processing strategy. This demonstration of the feasibility presents new opportunities for time-resolved experiments with microcrystals to study structural changes in real time at pink-beam synchrotron beamlines worldwide.

AB - Since the first successful serial crystallography (SX) experiment at a synchrotron radiation source, the popularity of this approach has continued to grow showing that third-generation synchrotrons can be viable alternatives to scarce X-ray free-electron laser sources. Synchrotron radiation flux may be increased ∼100 times by a moderate increase in the bandwidth ('pink beam' conditions) at some cost to data analysis complexity. Here, we report the first high-viscosity injector-based pink-beam SX experiments. The structures of proteinase K (PK) and A 2A adenosine receptor (A 2A AR) were determined to resolutions of 1.8 and 4.2Å using 4 and 24 consecutive 100ps X-ray pulse exposures, respectively. Strong PK data were processed using existing Laue approaches, while weaker A 2A AR data required an alternative data-processing strategy. This demonstration of the feasibility presents new opportunities for time-resolved experiments with microcrystals to study structural changes in real time at pink-beam synchrotron beamlines worldwide.

KW - X-ray crystallography

KW - injector-based serial crystallography

KW - membrane proteins

KW - pink-beam serial crystallography

KW - protein structures

KW - structural biology

KW - structure determination

KW - third-generation synchrotrons

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U2 - 10.1107/S205225251900263X

DO - 10.1107/S205225251900263X

M3 - Article

AN - SCOPUS:85065335933

SN - 2052-2525

VL - 6

SP - 412

EP - 425

JO - IUCrJ

JF - IUCrJ

ER -

High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source

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Keywords

ASJC Scopus subject areas

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