7 Å resolution in protein twodimensional-crystal X-ray diffraction at Linac Coherent Light Source

Bill Pedrini, Ching Ju Tsai, Guido Capitani, Celestino Padeste, Mark S. Hunter, Nadia Zatsepin, Anton Barty, W. Henry Benner, Sébastien Boutet, Geoffrey K. Feld, Stefan P. Hau-Riege, Richard Kirian, Christopher Kupitz, Marc Messerschmitt, John I. Ogren, Tommaso Pardini, Brent Segelke, Garth J. Williams, John Spence, Rafael AbelaMatthew Coleman, James E. Evans, Gebhard F X Schertler, Matthias Frank, Xiao Dan Li

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Membrane proteins arranged as two-dimensional crystals in the lipid environment provide close-to-physiological structural information, which is essential for understanding the molecular mechanisms of protein function. Previously, X-ray diffraction from individual two-dimensional crystals did not represent a suitable investigational tool because of radiation damage. The recent availability of ultrashort pulses from X-ray free-electron lasers (XFELs) has now provided a means to outrun the damage. Here, we report on measurements performed at the Linac Coherent Light Source XFEL on bacteriorhodopsin two-dimensional crystals mounted on a solid support and kept at room temperature. By merging data from about a dozen single crystal diffraction images, we unambiguously identified the diffraction peaks to a resolution of 7 A, thus improving the observable resolution with respect to that achievable from a single pattern alone. This indicates that a larger dataset will allow for reliable quantification of peak intensities, and in turn a corresponding increase in the resolution. The presented results pave the way for further XFEL studies on two-dimensional crystals, which may include pump-probe experiments at subpicosecond time resolution.

Original languageEnglish (US)
Article number20130500
JournalPhilosophical Transactions of the Royal Society B: Biological Sciences
Volume369
Issue number1647
DOIs
Publication statusPublished - Jul 17 2014

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Keywords

  • Bacteriorhodopsin
  • Crystallographic data analysis
  • Two-dimensional protein crystal
  • X-ray diffraction
  • X-ray free-electron laser

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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