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Structural and Functional Characterization of Schistosoma mansoni Cathepsin B1
A. Jílková, M. Horn, M. Mareš
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Enzyme Activation MeSH
- Electroporation MeSH
- Gene Expression MeSH
- Genetic Vectors metabolism MeSH
- Glycosylation MeSH
- Cathepsin B antagonists & inhibitors chemistry isolation & purification metabolism MeSH
- Kinetics MeSH
- Crystallization MeSH
- Mutation genetics MeSH
- Recombinant Proteins isolation & purification metabolism MeSH
- Saccharomycetales genetics MeSH
- Schistosoma mansoni enzymology MeSH
- Transformation, Genetic MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
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.
References provided by Crossref.org
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