title: In vivo crystallography at X-ray free-electron lasers: the next generation of structural biology?
abstract: The serendipitous discovery of the spontaneous growth of protein crystals inside cells has opened the field of crystallography to chemically unmodified samples directly available from their natural environment. On the one hand, through in vivo crystallography, protocols for protein crystal preparation can be highly simplified, although the technique suffers from difficulties in sampling, particularly in the extraction of the crystals from the cells partly due to their small sizes. On the other hand, the extremely intense X-ray pulses emerging from X-ray free-electron laser (XFEL) sources, along with the appearance of serial femtosecond crystallography (SFX) is a milestone for radiation damage-free protein structural studies but requires micrometre-size crystals. The combination of SFX with in vivo crystallography has the potential to boost the applicability of these techniques, eventually bringing the field to the point where in vitro sample manipulations will no longer be required, and direct imaging of the crystals from within the cells will be achievable. To fully appreciate the diverse aspects of sample characterization, handling and analysis, SFX experiments at the Japanese SPring-8 angstrom compact free-electron laser were scheduled on various types of in vivo grown crystals. The first experiments have demonstrated the feasibility of the approach and suggest that future in vivo crystallography applications at XFELs will be another alternative to nano-crystallography.
authors.affiliation: Photon Factory, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba, Ibaraki 305-0801, Japan.
authors.forename: François-Xavier
authors.initials: FX
authors.lastname: Gallat
authors.affiliation: Photon Factory, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba, Ibaraki 305-0801, Japan.
authors.forename: Naohiro
authors.initials: N
authors.lastname: Matsugaki
authors.affiliation: National Center for Advancing Translational Sciences, National Institutes of Health, 9800 Medical Center Drive, Rockville, MD 20850, USA.
authors.forename: Nathan P
authors.initials: NP
authors.lastname: Coussens
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, Canada M5S 3G5.
authors.forename: Koichiro J
authors.initials: KJ
authors.lastname: Yagi
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Building 76, Clayton, Victoria 3800, Australia.
authors.forename: Marion
authors.initials: M
authors.lastname: Boudes
authors.affiliation: Department of Medicinal Biotechnology, University of Tokushima, 1-78 Sho-machi Tokushima, Tokushima 770-8505, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Higashi
authors.affiliation: Department of Medicinal Biotechnology, University of Tokushima, 1-78 Sho-machi Tokushima, Tokushima 770-8505, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Tsuji
authors.affiliation: Deparment of Microbiology and Immunology, School of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Tatano
authors.affiliation: Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Division of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Eiichi
authors.initials: E
authors.lastname: Mizohata
authors.affiliation: Japan Synchrotron Radiation Research Institute, Kouto 1-1-1, Sayo, Hyogo 679-5198, Japan.
authors.forename: Kensuke
authors.initials: K
authors.lastname: Tono
authors.affiliation: Japan Synchrotron Radiation Research Institute, Kouto 1-1-1, Sayo, Hyogo 679-5198, Japan.
authors.forename: Yasumasa
authors.initials: Y
authors.lastname: Joti
authors.affiliation: Japan Synchrotron Radiation Research Institute, Kouto 1-1-1, Sayo, Hyogo 679-5198, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Kameshima
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Jaehyun
authors.initials: J
authors.lastname: Park
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Changyong
authors.initials: C
authors.lastname: Song
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Takaki
authors.initials: T
authors.lastname: Hatsui
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Makina
authors.initials: M
authors.lastname: Yabashi
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Nango
authors.affiliation: Department of Medicinal Biotechnology, University of Tokushima, 1-78 Sho-machi Tokushima, Tokushima 770-8505, Japan.
authors.forename: Kohji
authors.initials: K
authors.lastname: Itoh
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Building 76, Clayton, Victoria 3800, Australia.
authors.forename: Fasséli
authors.initials: F
authors.lastname: Coulibaly
authors.affiliation: Department of Cell and Systems Biology, University of Toronto, Toronto, Canada M5S 3G5.
authors.forename: Stephen
authors.initials: S
authors.lastname: Tobe
authors.affiliation: Institute for Stem Cell Biology and Regenerative Medicine, Bellary Road, Bangalore 560065, India.
authors.forename: S
authors.initials: S
authors.lastname: Ramaswamy
authors.affiliation: Department of Biology, University of Iowa, Iowa City, IA 52242, USA.
authors.forename: Barbara
authors.initials: B
authors.lastname: Stay
authors.affiliation: RIKEN SPring-8 Center, Kouto 1-1-1, Sayo, Hyogo 679-5148, Japan s.iwata@mfour.med.kyoto-u.ac.jp.
authors.forename: So
authors.initials: S
authors.lastname: Iwata
authors.affiliation: Photon Factory, High Energy Accelerator Research Organization, 1-1 Oho, Tsukuba, Ibaraki 305-0801, Japan Center for Free-Electron Laser science, Notkestrasse 85, Building 99, Hamburg 22607, Germany leonard.chavas@desy.de.
authors.forename: Leonard M G
authors.initials: LM
authors.lastname: Chavas
mesh_terms: D000818:Animals; D001695:Biology / Q000379:methods* / Q000639:trends; D016466:CHO Cells; D003058:Cockroaches; D006224:Cricetinae; D003412:Cricetulus; D018360:Crystallography, X-Ray / Q000379:methods*; D004583:Electrons; D006801:Humans; D007834:Lasers; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D011506:Proteins / Q000737:chemistry* / Q000648:ultrastructure; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D011506:Proteins
keywords: X-ray free-electron laser; in vivo crystallography; serial femtosecond crystallography