Schistosoma mansoni: Methods and Protocols

0525103 - ÚOCHB 2021 RIV US eng M - Část monografie knihy
Jílková, Adéla - Horn, Martin - Mareš, Michael
Structural and Functional Characterization of Schistosoma mansoni Cathepsin B1.
Schistosoma mansoni: Methods and Protocols. New York: Humana Press, 2020 - (Timson, D.), s. 145-158. Methods in Molecular Biology, 2151. ISBN 978-1-0716-0634-6
Grant CEP: GA MZd(CZ) NV18-05-00345; GA MŠMT LO1302
Institucionální podpora: RVO:61388963
Klíčová slova: parasite * Schistosoma mansoni * protease * cathepsin B * expression * activity assay * inhibition * crystal structure
Obor OECD: Biochemistry and molecular biology

Schistosomiasis caused by parasitic blood flukes of the genus Schistosoma is a global health problem with over 200 million people infected. Schistosoma mansoni cathepsin B1 (SmCB1) is a gut-associated protease critical for digestion of host blood proteins as a source of nutrients. SmCB1 is a validated drug target, and inhibitors of SmCB1 represent promising anti-schistosomals. A comprehensive structural and functional characterization of SmCB1 provides a starting point for the rational design of selective and potent SmCB1 inhibitors. Here, we report optimized protocols for (1) the production of recombinant SmCB1 in the Pichia pastoris expression system and its purification, (2) the measurement of SmCB1 activity and inhibition in a kinetic fluorescence assay, and (3) the preparation and crystallization of SmCB1 in complex with a model vinyl sulfone inhibitor, and the determination of its crystal structure.
Trvalý link: http://hdl.handle.net/11104/0309316