Towards time-resolved serial crystallography in a microfluidic device
Category
Published on
Type
journal-article
Author
Ashtamurthy S. Pawate and Vukica Šrajer and Jeremy Schieferstein and Sudipto Guha and Robert Henning and Irina Kosheleva and Marius Schmidt and Zhong Ren and Paul J. A. Kenis and Sarah L. Perry
Citation
Pawate, A.S. et al., 2015. Towards time-resolved serial crystallography in a microfluidic device. Acta Crystallogr Sect F Struct Biol Cryst Commun, 71(7), pp.823–830. Available at: http://dx.doi.org/10.1107/s2053230x15009061.
Abstract
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 achievedviapulsed laser illumination, and the resultant electron-density difference maps clearly depict the expected pR1/pRE46Qand pR2/pRCWstates at 10 µs and the pB1intermediate 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.
DOI
Funding
NSF-STC Biology with X-ray Lasers (NSF-1231306)
