Previous proof-of-concept measurements on single-layer two-dimensional membrane-protein crystals performed at X-ray free-electron lasers (FELs) have demonstrated that the collection of meaningful diffraction patterns, which is not possible at synchrotrons because of radiation-damage issues, is feasible. Here, the results obtained from the analysis of a thousand single-shot, room-temperature X-ray FEL diffraction images from two-dimensional crystals of a bacteriorhodopsin mutant are reported in detail. The high redundancy in the measurements boosts the intensity signal-to-noise ratio, so that the values of the diffracted intensities can be reliably determined down to the detector-edge resolution of 4 Å. The results show that two-dimensional serial crystallography at X-ray FELs is a suitable method to study membrane proteins to near-atomic length scales at ambient temperature. The method presented here can be extended to pump–probe studies of optically triggered structural changes on submillisecond timescales in two-dimensional crystals, which allow functionally relevant large-scale motions that may be quenched in three-dimensional crystals.
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- Resolution Extension by Image Summing in Serial Femtosecond Crystallography of Two-Dimensional Membrane-Protein Crystals
- Casadei, Cecilia M. (Author)
- Tsai, Ching-Ju (Author)
- Barty, Anton (Author)
- Hunter, Mark S. (Author)
- Zatsepin, Nadia (Author)
- Padeste, Celestino (Author)
- Capitani, Guido (Author)
- Benner, W. Henry (Author)
- Boutet, Sebastien (Author)
- Hau-Riege, Stefan P. (Author)
- Kupitz, Christopher (Author)
- Messerschmidt, Marc (Author)
- Ogren, John I. (Author)
- Pardini, Tom (Author)
- Rothschild, Kenneth J. (Author)
- Sala, Leonardo (Author)
- Segelke, Brent (Author)
- Williams, Garth J. (Author)
- Evans, James E. (Author)
- Li, Xiao-Dan (Author)
- Coleman, Matthew (Author)
- Pedrini, Bill (Author)
- Frank, Matthias (Author)
- College of Liberal Arts and Sciences (Contributor)
- Digital object identifier: 10.1107/S2052252517017043
- Identifier TypeInternational standard serial numberIdentifier Value2052-2525
- View the article as published at http://journals.iucr.org/m/issues/2018/01/00/ec5006/index.html
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Casadei, C. M., Tsai, C., Barty, A., Hunter, M. S., Zatsepin, N. A., Padeste, C., . . . Frank, M. (2018). Resolution extension by image summing in serial femtosecond crystallography of two-dimensional membrane-protein crystals. IUCrJ, 5(1), 103-117. doi:10.1107/s2052252517017043