title: Towards time-resolved serial crystallography in a microfluidic device.
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 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 µs and the pB1 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.
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Ashtamurthy S
authors.initials: AS
authors.lastname: Pawate
authors.affiliation: Center for Advanced Radiation Sources, The University of Chicago, Argonne, Illinois, USA.
authors.forename: Vukica
authors.initials: V
authors.lastname: Šrajer
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Schieferstein
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Sudipto
authors.initials: S
authors.lastname: Guha
authors.affiliation: Center for Advanced Radiation Sources, The University of Chicago, Argonne, Illinois, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Henning
authors.affiliation: Center for Advanced Radiation Sources, The University of Chicago, Argonne, Illinois, USA.
authors.forename: Irina
authors.initials: I
authors.lastname: Kosheleva
authors.affiliation: Department of Physics, The University of Wisconsin Milwaukee, Milwaukee, Wisconsin, USA.
authors.forename: Marius
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Center for Advanced Radiation Sources, The University of Chicago, Argonne, Illinois, USA.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Ren
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Paul J A
authors.initials: PJ
authors.lastname: Kenis
authors.affiliation: Department of Chemical and Biomolecular Engineering, The University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Perry
mesh_terms: D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry*; D018360:Crystallography, X-Ray / Q000379:methods; D056656:Lab-On-A-Chip Devices; D020283:Photoreceptors, Microbial / Q000032:analysis / Q000737:chemistry*; D013997:Time Factors; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D020283:Photoreceptors, Microbial; C050162:photoactive yellow protein, Bacteria
keywords: Laue diffraction; microfluidics; photoactive yellow protein; protein crystallization; serial crystallography; time-resolved protein crystallography