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 Spence, Petra Fromme, Nadia Zatsepin, Robert F. Fischetti, Vadim Cherezov & 1 others Wei Liu

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 languageEnglish (US)
Pages (from-to)412-425
Number of pages14
JournalIUCrJ
Volume6
DOIs
StatePublished - Jan 1 2019

Fingerprint

Crystallography
Microcrystals
Synchrotrons
microcrystals
Synchrotron radiation
radiation sources
injectors
Viscosity
Adenosine A2A Receptors
crystallography
adenosines
Endopeptidase K
synchrotron radiation
Radiation
viscosity
synchrotrons
X ray lasers
Experiments
Free electron lasers
X-Rays

Keywords

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

ASJC Scopus subject areas

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

Cite this

High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source. / Martin-Garcia, Jose M.; Zhu, Lan; Mendez, Derek; Lee, Ming Yue; Chun, Eugene; Li, Chufeng; Hu, Hao; Subramanian, Ganesh; Kissick, David; Ogata, Craig; Henning, Robert; Ishchenko, Andrii; Dobson, Zachary; Zhang, Shangji; Weierstall, Uwe; Spence, John; Fromme, Petra; Zatsepin, Nadia; Fischetti, Robert F.; Cherezov, Vadim; Liu, Wei.

In: IUCrJ, Vol. 6, 01.01.2019, p. 412-425.

Research output: Contribution to journalArticle

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, J, Fromme, P, Zatsepin, N, 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
Martin-Garcia, Jose M. ; Zhu, Lan ; Mendez, Derek ; Lee, Ming Yue ; Chun, Eugene ; Li, Chufeng ; Hu, Hao ; Subramanian, Ganesh ; Kissick, David ; Ogata, Craig ; Henning, Robert ; Ishchenko, Andrii ; Dobson, Zachary ; Zhang, Shangji ; Weierstall, Uwe ; Spence, John ; Fromme, Petra ; Zatsepin, Nadia ; Fischetti, Robert F. ; Cherezov, Vadim ; Liu, Wei. / High-viscosity injector-based pink-beam serial crystallography of microcrystals at a synchrotron radiation source. In: IUCrJ. 2019 ; Vol. 6. pp. 412-425.
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