Expression of an active cathepsin B-like protein Sm31 from Schistosoma mansoni in insect cells.
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.
Götz
B
B
Institute of Parasitology, University of Zürich, Switzerland.
Felleisen
R
R
Shaw
E
E
Klinkert
M Q
MQ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002401:Cathepsin B / Q000096:biosynthesis* / Q000235:genetics; D002478:Cells, Cultured; D005822:Genetic Vectors; D009036:Moths / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012550:Schistosoma mansoni / Q000201:enzymology* / Q000235:genetics
D011994:Recombinant Proteins; D002401:Cathepsin B