Abstract
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.
Original language | English (US) |
---|---|
Pages (from-to) | 103-117 |
Number of pages | 15 |
Journal | IUCrJ |
Volume | 5 |
DOIs | |
State | Published - Jan 1 2018 |
Externally published | Yes |
Keywords
- free-electron lasers
- membrane proteins
- serial crystallography
- two-dimensional crystals
ASJC Scopus subject areas
- Chemistry(all)
- Biochemistry
- Materials Science(all)
- Condensed Matter Physics