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Expression of an active cathepsin B-like protein Sm31 from Schistosoma mansoni in insect cells
1Institute of Parasitology, University of Zürich, Switzerland.
Tropical Medicine and Parasitology : Official Organ of Deutsche Tropenmedizinische Gesellschaft and of Deutsche Gesellschaft Fur Technische Zusammenarbeit (GTZ)
|December 1, 1992
Abstract:
Recombinant Schistosoma mansoni cathepsin B was produced in insect cells via the baculovirus expression vector system as a 37.5 kDa precursor molecule and a 31 kDa mature enzyme. Extracts prepared from cells infected with the recombinant virus were able to cleave a synthetic dipeptide substrate specific for cathepsin B. Proteolytic activity was inhibited by trans-epoxysuccinyl-L-leucyl-amido (4-guanidino) butane (E64) but not by phenylmethylsulphonyl-fluoride (PMSF), pepstatin and 1,10-phenanthroline. Specific inhibition by diazomethylketone derivatives, which bind covalently to the active centre of cysteinyl proteinases was also demonstrated.