title: Time-resolved serial crystallography captures high-resolution intermediates of photoactive yellow protein.
abstract: Serial femtosecond crystallography using ultrashort pulses from x-ray free electron lasers (XFELs) enables studies of the light-triggered dynamics of biomolecules. We used microcrystals of photoactive yellow protein (a bacterial blue light photoreceptor) as a model system and obtained high-resolution, time-resolved difference electron density maps of excellent quality with strong features; these allowed the determination of structures of reaction intermediates to a resolution of 1.6 angstroms. Our results open the way to the study of reversible and nonreversible biological reactions on time scales as short as femtoseconds under conditions that maximize the extent of reaction initiation throughout the crystal.
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Tenboer
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Shibom
authors.initials: S
authors.lastname: Basu
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Nadia
authors.initials: N
authors.lastname: Zatsepin
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.
authors.forename: Kanupriya
authors.initials: K
authors.lastname: Pande
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Despina
authors.initials: D
authors.lastname: Milathianaki
authors.affiliation: Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Frank
authors.affiliation: Lawrence Livermore National Laboratory, Livermore, CA 94550, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Hunter
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Jason E
authors.initials: JE
authors.lastname: Koglin
authors.affiliation: Centre for Ultrafast Imaging, University of Hamburg, 22761 Hamburg, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: Oberthuer
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Heymann
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Kupitz
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Chelsie
authors.initials: C
authors.lastname: Conrad
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Coe
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Shatabdi
authors.initials: S
authors.lastname: Roy-Chowdhury
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Uwe
authors.initials: U
authors.lastname: Weierstall
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: James
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Dingjie
authors.initials: D
authors.lastname: Wang
authors.affiliation: Hauptman-Woodward Institute, State University of New York at Buffalo, 700 Ellicott Street, Buffalo, NY 14203, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Grant
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Center for Free Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Oleksandr
authors.initials: O
authors.lastname: Yefanov
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Scales
authors.affiliation: Centre for Ultrafast Imaging, University of Hamburg, 22761 Hamburg, Germany.|Center for Free Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Cornelius
authors.initials: C
authors.lastname: Gati
authors.affiliation: Centre for Ultrafast Imaging, University of Hamburg, 22761 Hamburg, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Seuring
authors.affiliation: Center for Advanced Radiation Sources, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Vukica
authors.initials: V
authors.lastname: Srajer
authors.affiliation: Center for Advanced Radiation Sources, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Henning
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Schwander
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Raimund
authors.initials: R
authors.lastname: Fromme
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.
authors.forename: Abbas
authors.initials: A
authors.lastname: Ourmazd
authors.affiliation: Center for Advanced Radiation Sources, University of Chicago, Chicago, IL 60637, USA.|Department of Biochemistry and Molecular Biology and Institute for Biophysical Dynamics, University of Chicago, Chicago, IL 60637, USA.
authors.forename: Keith
authors.initials: K
authors.lastname: Moffat
authors.affiliation: Faculty of Natural Sciences, Life Sciences, Imperial College, London SW7 2AZ, UK.
authors.forename: Jasper J
authors.initials: JJ
authors.lastname: Van Thor
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: John C H
authors.initials: JC
authors.lastname: Spence
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Petra
authors.initials: P
authors.lastname: Fromme
authors.affiliation: Centre for Ultrafast Imaging, University of Hamburg, 22761 Hamburg, Germany.|Center for Free Electron Laser Science, Deutsches Elektronen Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Henry N
authors.initials: HN
authors.lastname: Chapman
authors.affiliation: Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA.|Physics Department, University of Wisconsin, Milwaukee, WI 53211, USA. m-schmidt@uwm.edu.
authors.forename: Marius
authors.initials: M
authors.lastname: Schmidt
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000648:ultrastructure*; D018360:Crystallography, X-Ray / Q000379:methods*; D020283:Photoreceptors, Microbial / Q000737:chemistry / Q000648:ultrastructure*; D011487:Protein Conformation; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D020283:Photoreceptors, Microbial; C050162:photoactive yellow protein, Bacteria