title: Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography.
abstract: Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3 Å resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1 ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1 mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Przemyslaw
authors.identifier: 0000-0002-8040-7753
authors.initials: P
authors.lastname: Nogly
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Valerie
authors.initials: V
authors.lastname: Panneels
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Garrett
authors.initials: G
authors.lastname: Nelson
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Cornelius
authors.initials: C
authors.lastname: Gati
authors.affiliation: Biometal Science Laboratory, RIKEN SPring-8 Center, Hyogo 679-5148, Japan.
authors.forename: Tetsunari
authors.initials: T
authors.lastname: Kimura
authors.affiliation: SwissFEL, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Christopher
authors.identifier: 0000-0003-4714-9139
authors.initials: C
authors.lastname: Milne
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Despina
authors.initials: D
authors.lastname: Milathianaki
authors.affiliation: Biometal Science Laboratory, RIKEN SPring-8 Center, Hyogo 679-5148, Japan.|PRESTO, JST, Saitama 332-0012, Japan.
authors.forename: Minoru
authors.initials: M
authors.lastname: Kubo
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Wenting
authors.initials: W
authors.lastname: Wu
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Chelsie
authors.initials: C
authors.lastname: Conrad
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Jesse
authors.initials: J
authors.lastname: Coe
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Bean
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, SE-40530 Gothenburg, Sweden.
authors.forename: Petra
authors.initials: P
authors.lastname: Båth
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, SE-40530 Gothenburg, Sweden.
authors.forename: Robert
authors.initials: R
authors.lastname: Dods
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, SE-40530 Gothenburg, Sweden.
authors.forename: Rajiv
authors.initials: R
authors.lastname: Harimoorthy
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Beyerlein
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Jan
authors.initials: J
authors.lastname: Rheinberger
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Daniel
authors.identifier: 0000-0002-8348-6661
authors.initials: D
authors.lastname: James
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: DePonte
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Chufeng
authors.initials: C
authors.lastname: Li
authors.affiliation: SwissFEL, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Sala
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Hunter
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Jason E
authors.identifier: 0000-0001-6811-6083
authors.initials: JE
authors.lastname: Koglin
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, SE-40530 Gothenburg, Sweden.
authors.forename: Peter
authors.initials: P
authors.lastname: Berntsen
authors.affiliation: SACLA Science Research Group, RIKEN/SPring-8 Center, Hyogo 679-5148, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Nango
authors.affiliation: SACLA Science Research Group, RIKEN/SPring-8 Center, Hyogo 679-5148, Japan.|Department of Cell Biology, Kyoto University, Kyoto 606-8501, Japan.
authors.forename: So
authors.initials: S
authors.lastname: Iwata
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, 22761 Hamburg, Germany.|Centre for Ultrafast Imaging, University of Hamburg, 22761 Hamburg, Germany.
authors.forename: Henry N
authors.identifier: 0000-0002-4655-1743
authors.initials: HN
authors.lastname: Chapman
authors.affiliation: Department of Chemistry and Biochemistry, and Center for Applied Structural Discovery, Biodesign Institute, Arizona State University, Tempe, Arizona 85287-1604, USA.
authors.forename: Petra
authors.initials: P
authors.lastname: Fromme
authors.affiliation: Lawrence Livermore National Laboratory, Livermore 94550, USA.
authors.forename: Matthias
authors.identifier: 0000-0002-5244-0791
authors.initials: M
authors.lastname: Frank
authors.affiliation: SwissFEL, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Rafael
authors.initials: R
authors.lastname: Abela
authors.affiliation: Linac Coherent Light Source (LCLS), SLAC National Accelerator Laboratory, Menlo Park, California 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, 22607 Hamburg, Germany.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: White
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: Uwe
authors.initials: U
authors.lastname: Weierstall
authors.affiliation: Department of Physics, Arizona State University, Tempe, Arizona 85287, USA.
authors.forename: John
authors.initials: J
authors.lastname: Spence
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, Box 462, SE-40530 Gothenburg, Sweden.
authors.forename: Richard
authors.initials: R
authors.lastname: Neutze
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.|Department of Biology, ETH Zurich, Zürich 8093, Switzerland.
authors.forename: Gebhard
authors.initials: G
authors.lastname: Schertler
authors.affiliation: Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.
authors.forename: Jörg
authors.initials: J
authors.lastname: Standfuss
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry*; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D005240:Feasibility Studies; D007834:Lasers; D008055:Lipids / Q000737:chemistry*; D011487:Protein Conformation; D017356:Synchrotrons; D013997:Time Factors; D014783:Viscosity; D056928:X-Ray Absorption Spectroscopy / Q000295:instrumentation / Q000379:methods
chemical_list: D008055:Lipids; D001436:Bacteriorhodopsins