Journal
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS
Volume 71, Issue -, Pages 823-830Publisher
INT UNION CRYSTALLOGRAPHY
DOI: 10.1107/S2053230X15009061
Keywords
serial crystallography; Laue diffraction; time-resolved protein crystallography; protein crystallization; microfluidics; photoactive yellow protein
Funding
- National Institutes of Health [GM086727]
- US Department of Energy Basic Energy Sciences Office of Science [DE-AC02-06CH11357]
- National Institutes of Health National Institute of General Medical Sciences [1R24GM111072]
- NFS-STC [1231306]
- [NSF-0952643]
- Div Of Molecular and Cellular Bioscience
- Direct For Biological Sciences [0952643] Funding Source: National Science Foundation
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Serial methods for crystallography have the potential to enable dynamic structural studies of protein targets that have been resistant to single-crystal strategies. The use of serial data-collection strategies can circumvent challenges associated with radiation damage and repeated reaction initiation. This work utilizes a microfluidic crystallization platform for the serial time-resolved Laue diffraction analysis of macroscopic crystals of photoactive yellow protein (PYP). Reaction initiation was achieved via pulsed laser illumination, and the resultant electron-density difference maps clearly depict the expected pR(1)/pR(E46Q) and pR(2)/pR(CW) states at 10 mu s and the pB(1) intermediate at 1 ms. The strategies presented here have tremendous potential for extension to chemical triggering methods for reaction initiation and for extension to dynamic, multivariable analyses.
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