title: Natively inhibited Trypanosoma brucei cathepsin B structure determined by using an X-ray laser.
abstract: The Trypanosoma brucei cysteine protease cathepsin B (TbCatB), which is involved in host protein degradation, is a promising target to develop new treatments against sleeping sickness, a fatal disease caused by this protozoan parasite. The structure of the mature, active form of TbCatB has so far not provided sufficient information for the design of a safe and specific drug against T. brucei. By combining two recent innovations, in vivo crystallization and serial femtosecond crystallography, we obtained the room-temperature 2.1 angstrom resolution structure of the fully glycosylated precursor complex of TbCatB. The structure reveals the mechanism of native TbCatB inhibition and demonstrates that new biomolecular information can be obtained by the "diffraction-before-destruction" approach of x-ray free-electron lasers from hundreds of thousands of individual microcrystals.
authors.affiliation: Joint Laboratory for Structural Biology of Infection and Inflammation, Institute of Biochemistry and Molecular Biology, University of Hamburg, and Institute of Biochemistry, University of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany.|German Centre for Infection Research, University of Lübeck, 23538 Lübeck, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Redecke
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Karol
authors.initials: K
authors.lastname: Nass
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Daniel P
authors.initials: DP
authors.lastname: DePonte
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: White
authors.affiliation: Joint Laboratory for Structural Biology of Infection and Inflammation, Institute of Biochemistry and Molecular Biology, University of Hamburg, and Institute of Biochemistry, University of Lübeck, at Deutsches Elektronen-Synchrotron (DESY), Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Dirk
authors.initials: D
authors.lastname: Rehders
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Barty
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Francesco
authors.initials: F
authors.lastname: Stellato
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Mengning
authors.initials: M
authors.lastname: Liang
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Thomas R M
authors.initials: TRM
authors.lastname: Barends
authors.affiliation: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Boutet
authors.affiliation: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Garth J
authors.initials: GJ
authors.lastname: Williams
authors.affiliation: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Marc
authors.initials: M
authors.lastname: Messerschmidt
authors.affiliation: Linac Coherent Light Source, Stanford Linear Accelerator Center (SLAC) National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: M Marvin
authors.initials: MM
authors.lastname: Seibert
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Andrew
authors.initials: A
authors.lastname: Aquila
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, SE-405 30 Gothenburg, Sweden.
authors.forename: David
authors.initials: D
authors.lastname: Arnlund
authors.affiliation: Photon Science, DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Sasa
authors.initials: S
authors.lastname: Bajt
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Barth
authors.affiliation: Photon Ultrafast Laser Science and Engineering (PULSE) Institute, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, CA 94025, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bogan
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Carl
authors.initials: C
authors.lastname: Caleman
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Tzu-Chiao
authors.initials: TC
authors.lastname: Chao
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: R Bruce
authors.initials: RB
authors.lastname: Doak
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Fleckenstein
authors.affiliation: Lawrence Livermore National Laboratory, 7000 East Avenue, Livermore, CA 94550, USA.
authors.forename: Matthias
authors.initials: M
authors.lastname: Frank
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: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Galli
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Ingo
authors.initials: I
authors.lastname: Grotjohann
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Hunter
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, SE-405 30 Gothenburg, Sweden.
authors.forename: Linda C
authors.initials: LC
authors.lastname: Johansson
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Kassemeyer
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, SE-405 30 Gothenburg, Sweden.
authors.forename: Gergely
authors.initials: G
authors.lastname: Katona
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.|Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Kirian
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Koopmann
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Chris
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authors.lastname: Kupitz
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Lukas
authors.initials: L
authors.lastname: Lomb
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Andrew V
authors.initials: AV
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authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Mogk
authors.affiliation: Department of Chemistry and Molecular Biology, University of Gothenburg, SE-405 30 Gothenburg, Sweden.
authors.forename: Richard
authors.initials: R
authors.lastname: Neutze
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Shoeman
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Steinbrener
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, Husargatan 3, SE-75124 Uppsala, Sweden.
authors.forename: Nicusor
authors.initials: N
authors.lastname: Timneanu
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Dingjie
authors.initials: D
authors.lastname: Wang
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: Nadia A
authors.initials: NA
authors.lastname: Zatsepin
authors.affiliation: Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: John C H
authors.initials: JCH
authors.lastname: Spence
authors.affiliation: Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Petra
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authors.lastname: Fromme
authors.affiliation: Max-Planck-Institut für medizinische Forschung, Jahnstrasse 29, 69120 Heidelberg, Germany.|Max Planck Advanced Study Group, Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Ilme
authors.initials: I
authors.lastname: Schlichting
authors.affiliation: Interfaculty Institute of Biochemistry, University of Tübingen, Hoppe-Seyler-Strasse 4, 72076 Tübingen, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Duszenko
authors.affiliation: Institute of Biochemistry and Molecular Biology, University of Hamburg, at DESY, Notkestrasse 85, 22607 Hamburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Betzel
authors.affiliation: Center for Free-Electron Laser Science (CFEL), DESY, Notkestrasse 85, 22607 Hamburg, Germany.|Department of Physics, University of Hamburg, Luruper Chaussee 149, 22761 Hamburg, Germany.
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mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D004792:Enzyme Precursors / Q000737:chemistry; D006031:Glycosylation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015800:Protozoan Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D061987:Sf9 Cells; D018411:Spodoptera; D014346:Trypanosoma brucei brucei / Q000201:enzymology*; D014965:X-Rays
chemical_list: D004792:Enzyme Precursors; D015800:Protozoan Proteins; D002401:Cathepsin B