Purification and characterization of a 47 kDa protease from Schistosoma mansoni cercarial secretion.
Fractionation of Schistosoma mansoni cercariae gland secretion on a Sephadex G-150 column followed by a Superose-12 column in an FPLC system, isolated a 47 kDa protease which migrated as a single band on SDS-PAGE gels. A monoclonal antibody (MAb) was produced which recognizes only the 47 kDa protease, and an immuno-affinity column with the MAb was used to isolate the protease. The 47 kDa protease showed activity on several macromolecules such as elastin and collagen type VI besides gelatin and casein. This suggests that this enzyme can be one of the enzymes that might facilitate invasion of the cercariae through host skin. The optimal pH of the protease against the synthetic substrate, Ac-Phe-Arg-Nan, in Tris-HCl buffer was 10. Experiments with protease inhibitors indicate that the purified enzyme is a serine protease.
Chavez-Olortegui
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Departamento de Bioquímica-Imunologia, Instituto de Ciências Biológicas da Universidade Federal de Minas Gerais, Brazil.
Resende
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Tavares
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D002364:Caseins / Q000378:metabolism; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003094:Collagen / Q000378:metabolism; D004549:Elastin / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D012550:Schistosoma mansoni / Q000201:enzymology*; D012697:Serine Endopeptidases / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D002364:Caseins; D015842:Serine Proteinase Inhibitors; D005780:Gelatin; D005340:Fibrinogen; D003094:Collagen; D004549:Elastin; D012697:Serine Endopeptidases