Purification and characterization of proteases secreted by transforming schistosomula of Schistosoma mansoni

Moshe Marikovsky, Zvi Fishelson, Ruth Arnon

Research output: Contribution to journalArticlepeer-review

Abstract

Schistosomula of Schistosoma mansoni which are mechanically transformed at 4°C and are then incubated at 37°C in defined medium spontaneously secrete two proteases, a major one of 28 kDa and a minor one of 60 kDa. These were purified by ion exchange chromatography on DEAE-cellulose and gel filtration on Ultrogel AcA 54 with yields of 33% and 29%, respectively. Both appeared as single bands by silver staining following sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis. The 28 kDa protease is a glycoprotein that has a pI of 11 or higher and an optimal activity around pH 9.0. It cleaves casein, gelatin and human C3 and C3b. It is metal-ion independent and is inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, soy-bean trypsin inhibitor, α1 antitrypsin, Zn2+ ions, sodium dodecyl sulphate and normal human serum. The 60 kDa protease is a glycoprotein with a pI of 9.2. It can also cleave casein and gelatin and its activity is inhibited by phenylmethanesulfonyl fluoride but not by diisopropyl fluorophosphate or sodium dodecyl sulphate. We suggest that these proteases may play a role during cercarial penetration of the skin and in shedding of the cercarial glycocalyx.

Original languageEnglish
Pages (from-to)45-54
Number of pages10
JournalMolecular and Biochemical Parasitology
Volume30
Issue number1
DOIs
StatePublished - Jul 1988
Externally publishedYes

Keywords

  • Concanavalin A
  • Schistosoma mansoni
  • Schistosomulum
  • Secreted enzyme
  • Serine protease

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