Natively Inhibited Trypanosoma brucei Cathepsin B Structure Determined by Using an X-ray Laser

• Published in: Science (American Association for the Advancement of Science) Vol. 339; no. 6116; pp. 227 - 230
• Main Authors: Redecke, Lars, Nass, Karol, Deponte, Daniel P., White, Thomas A., Rehders, Dirk, Barty, Anton, Stellato, Francesco, Liang, Mengning, Barends, Thomas R.M., Boutet, Sébastien, Williams, Garth J., Messerschmidt, Marc, Seibert, M. Marvin, Aquila, Andrew, Arnlund, David, Bajt, Sasa, Barth, Torsten, Bogan, Michael J., Caleman, Carl, Chao, Tzu-Chiao, Doak, R. Bruce, Fleckenstein, Holger, Frank, Matthias, Fromme, Raimund, Galli, Lorenzo, Grotjohann, Ingo, Hunter, Mark S., Johansson, Linda C., Kassemeyer, Stephan, Katona, Gergely, Kirian, Richard A., Koopmann, Rudolf, Kupitz, Chris, Lomb, Lukas, Martin, Andrew V., Mogk, Stefan, Neutze, Richard, Shoeman, Robert L., Steinbrener, Jan, Timneanu, Nicusor, Wang, Dingjie, Weierstall, Uwe, Zatsepin, Nadia A., Spence, John C. H., Fromme, Petra, Schlichting, Ilme, Duszenko, Michael, Betzel, Christian, Chapman, Henry N.
• Format: Journal Article
• Language: English
• Published: United States American Association for the Advancement of Science 11-01-2013; The American Association for the Advancement of Science
• Subjects: Activation; Active sites; Amino Acid Sequence; Animals; antagonists & inhibitors; Atoms; Biochemistry; Biochemistry and Molecular Biology; Biokemi och molekylärbiologi; Catalytic Domain; Cathepsin B; Cathepsin B - antagonists & inhibitors; Cathepsin B - chemistry; chemistry; Crystal structure; Crystallization; Crystallography; Crystallography, X-Ray; Crystals; Diffraction; Electron density; Electronic structure; Enzyme Precursors; Enzyme Precursors - chemistry; Enzymes; enzymology; Glycosylation; Inhibitors; Lasers; Models, Molecular; Molecular; Molecular biology; Molecular Sequence Data; NATURAL SCIENCES; NATURVETENSKAP; Parasitic diseases; Proteases; Protein Conformation; Protozoa; Protozoan Proteins; Protozoan Proteins - antagonists & inhibitors; Protozoan Proteins - chemistry; Radiation damage; Sf9 Cells; Spodoptera; Trypanosoma brucei; Trypanosoma brucei brucei; Trypanosoma brucei brucei - enzymology; X-Ray; X-ray lasers; X-Rays
• ISSN: 0036-8075; 1095-9203; 1095-9203
• DOI: 10.1126/science.1229663

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.