title: Protein crystal structure obtained at 2.9 Å resolution from injecting bacterial cells into an X-ray free-electron laser beam.
abstract: It has long been known that toxins produced by Bacillus thuringiensis (Bt) are stored in the bacterial cells in crystalline form. Here we describe the structure determination of the Cry3A toxin found naturally crystallized within Bt cells. When whole Bt cells were streamed into an X-ray free-electron laser beam we found that scattering from other cell components did not obscure diffraction from the crystals. The resolution limits of the best diffraction images collected from cells were the same as from isolated crystals. The integrity of the cells at the moment of diffraction is unclear; however, given the short time (∼ 5 µs) between exiting the injector to intersecting with the X-ray beam, our result is a 2.9-Å-resolution structure of a crystalline protein as it exists in a living cell. The study suggests that authentic in vivo diffraction studies can produce atomic-level structural information.
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Sawaya
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and.
authors.forename: Duilio
authors.initials: D
authors.lastname: Cascio
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and.
authors.forename: Mari
authors.initials: M
authors.lastname: Gingery
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and.
authors.forename: Jose
authors.initials: J
authors.lastname: Rodriguez
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and.
authors.forename: Lukasz
authors.initials: L
authors.lastname: Goldschmidt
authors.affiliation: Université Grenoble Alpes, Centre National de la Recherche Scientifique, and Commissariat à l'Energie Atomique, Institut de Biologie Structurale, F-38044 Grenoble, France;
authors.forename: Jacques-Philippe
authors.initials: JP
authors.lastname: Colletier
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025;
authors.forename: Marc M
authors.initials: MM
authors.lastname: Messerschmidt
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025;
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025;
authors.forename: Jason E
authors.initials: JE
authors.lastname: Koglin
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025;
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720;
authors.forename: Aaron S
authors.initials: AS
authors.lastname: Brewster
authors.affiliation: Max Planck Institute for Medical Research, 69120 Heidelberg, Germany;
authors.forename: Karol
authors.initials: K
authors.lastname: Nass
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720;
authors.forename: Johan
authors.initials: J
authors.lastname: Hattne
authors.affiliation: Max Planck Institute for Medical Research, 69120 Heidelberg, Germany;
authors.forename: Sabine
authors.initials: S
authors.lastname: Botha
authors.affiliation: Max Planck Institute for Medical Research, 69120 Heidelberg, Germany; Department of Physics, Arizona State University, Tempe, AZ 85287; and.
authors.forename: R Bruce
authors.initials: RB
authors.lastname: Doak
authors.affiliation: Max Planck Institute for Medical Research, 69120 Heidelberg, Germany;
authors.forename: Robert L
authors.initials: RL
authors.lastname: Shoeman
authors.affiliation: Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, CA 94025;
authors.forename: Daniel P
authors.initials: DP
authors.lastname: DePonte
authors.affiliation: Department of Entomology and.
authors.forename: Hyun-Woo
authors.initials: HW
authors.lastname: Park
authors.affiliation: Department of Entomology and Graduate Program in Cell, Molecular and Developmental Biology, University of California, Riverside, CA 92521.
authors.forename: Brian A
authors.initials: BA
authors.lastname: Federici
authors.affiliation: Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720;
authors.forename: Nicholas K
authors.initials: NK
authors.lastname: Sauter
authors.affiliation: Max Planck Institute for Medical Research, 69120 Heidelberg, Germany;
authors.forename: Ilme
authors.initials: I
authors.lastname: Schlichting
authors.affiliation: UCLA-DOE Institute for Genomics and Proteomics, Department of Biological Chemistry, and Howard Hughes Medical Institute, University of California, Los Angeles, CA 90095-1570; david@mbi.ucla.edu.
authors.forename: David S
authors.initials: DS
authors.lastname: Eisenberg
mesh_terms: D001413:Bacillus thuringiensis / Q000737:chemistry* / Q000648:ultrastructure; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000295:instrumentation / Q000379:methods*; D004731:Endotoxins / Q000737:chemistry*; D006460:Hemolysin Proteins / Q000737:chemistry*; D007834:Lasers; D013171:Spores, Bacterial / Q000737:chemistry* / Q000648:ultrastructure; D017356:Synchrotrons; D014961:X-Ray Diffraction
chemical_list: D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis
keywords: Cry3A insecticidal toxin; XFEL; serial femtosecond crystallography