title: The room temperature crystal structure of a bacterial phytochrome determined by serial femtosecond crystallography.
abstract: Phytochromes are a family of photoreceptors that control light responses of plants, fungi and bacteria. A sequence of structural changes, which is not yet fully understood, leads to activation of an output domain. Time-resolved serial femtosecond crystallography (SFX) can potentially shine light on these conformational changes. Here we report the room temperature crystal structure of the chromophore-binding domains of the Deinococcus radiodurans phytochrome at 2.1 Å resolution. The structure was obtained by serial femtosecond X-ray crystallography from microcrystals at an X-ray free electron laser. We find overall good agreement compared to a crystal structure at 1.35 Å resolution derived from conventional crystallography at cryogenic temperatures, which we also report here. The thioether linkage between chromophore and protein is subject to positional ambiguity at the synchrotron, but is fully resolved with SFX. The study paves the way for time-resolved structural investigations of the phytochrome photocycle with time-resolved SFX.
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Petra
authors.initials: P
authors.lastname: Edlund
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.|Department of Biological and Environmental Sciences, University of Jyvaskyla, 40014 Jyvaskyla, Finland.|Faculty of Medicine, Anatomy, University of Helsinki, 00014 Helsinki, Finland.
authors.forename: Heikki
authors.initials: H
authors.lastname: Takala
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Elin
authors.initials: E
authors.lastname: Claesson
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Léocadie
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authors.lastname: Henry
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Robert
authors.initials: R
authors.lastname: Dods
authors.affiliation: Nanoscience Center, Department of Physics, University of Jyvaskyla, 40014 Jyvaskyla, Finland.
authors.forename: Heli
authors.initials: H
authors.lastname: Lehtivuori
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Matthijs
authors.initials: M
authors.lastname: Panman
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, D-22603 Hamburg, Germany.
authors.forename: Kanupriya
authors.initials: K
authors.lastname: Pande
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, D-22603 Hamburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: White
authors.affiliation: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, 113-0033, Tokyo, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Nakane
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Oskar
authors.initials: O
authors.lastname: Berntsson
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Emil
authors.initials: E
authors.lastname: Gustavsson
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Petra
authors.initials: P
authors.lastname: Båth
authors.affiliation: Nanoscience Center, Department of Chemistry, University of Jyvaskyla, 40014 Jyvaskyla, Finland.
authors.forename: Vaibhav
authors.initials: V
authors.lastname: Modi
authors.affiliation: School of Molecular Sciences, Arizona State University, 85287 Tempe, Arizona, USA.
authors.forename: Shatabdi
authors.initials: S
authors.lastname: Roy-Chowdhury
authors.affiliation: School of Molecular Sciences, Arizona State University, 85287 Tempe, Arizona, USA.
authors.forename: James
authors.initials: J
authors.lastname: Zook
authors.affiliation: Australian Research Council Centre of Excellence for Advanced Molecular Imaging, La Trobe Institute for Molecular Science, La Trobe University, 3086 Melbourne, Victoria, Australia.
authors.forename: Peter
authors.initials: P
authors.lastname: Berntsen
authors.affiliation: Physics Department, University of Wisconsin, WI 53211 Milwaukee, USA.
authors.forename: Suraj
authors.initials: S
authors.lastname: Pandey
authors.affiliation: Physics Department, University of Wisconsin, WI 53211 Milwaukee, USA.
authors.forename: Ishwor
authors.initials: I
authors.lastname: Poudyal
authors.affiliation: Physics Department, University of Wisconsin, WI 53211 Milwaukee, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Tenboer
authors.affiliation: Physics Department, University of Wisconsin, WI 53211 Milwaukee, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Kupitz
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, D-22603 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: School of Molecular Sciences, Arizona State University, 85287 Tempe, Arizona, USA.
authors.forename: Petra
authors.initials: P
authors.lastname: Fromme
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Jake D
authors.initials: JD
authors.lastname: Koralek
authors.affiliation: RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, 679-5148 Hyogo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Biodesign Institute, Arizona State University, 85287 Tempe, Arizona, USA.|Center for Applied Structural Discovery, Arizona State University, 85287 Tempe, Arizona, USA.|Department of Physics, Arizona State University, 85287 Tempe, Arizona, USA.
authors.forename: John
authors.initials: J
authors.lastname: Spence
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Mengning
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authors.lastname: Liang
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Hunter
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Sebastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, 679-5148 Hyogo, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Nango
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Chicago, 60637 Chicago, Illinois, USA.|BioCARS, Argonne National Laboratory, 60439 Argonne, Illinois USA.
authors.forename: Keith
authors.initials: K
authors.lastname: Moffat
authors.affiliation: Nanoscience Center, Department of Chemistry, University of Jyvaskyla, 40014 Jyvaskyla, Finland.
authors.forename: Gerrit
authors.initials: G
authors.lastname: Groenhof
authors.affiliation: Nanoscience Center, Department of Biological and Environmental Sciences, University of Jyvaskyla, 40014 Jyvaskyla, Finland.
authors.forename: Janne
authors.initials: J
authors.lastname: Ihalainen
authors.affiliation: Department of Biology, Northeastern Illinois University, 5500 North St. Louis Avenue, 60625 Chicago, Illinois, United States.
authors.forename: Emina A
authors.initials: EA
authors.lastname: Stojković
authors.affiliation: Physics Department, University of Wisconsin, WI 53211 Milwaukee, USA.
authors.forename: Marius
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, 40530 Gothenburg, Sweden.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Westenhoff
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D034301:Deinococcus / Q000737:chemistry*; D010834:Phytochrome / Q000737:chemistry*; D011487:Protein Conformation; D013696:Temperature
chemical_list: D010834:Phytochrome