Isolation of a cDNA encoding Fasciola hepatica cathepsin L2 and functional expression in Saccharomyces cerevisiae

摘要: Abstract Cathepsin L2 is a major cysteine proteinase secreted by adult Fasciola hepatica . The enzyme differs from other reported cathepsin Ls in that it can cleave peptide substrates contain proline the P 2 position. A cDNA was isolated an expression library immunoscreening with antiserum prepared against purified native L2. This sequenced and shown to encode complete preprocathepsin L proteinase. Functionally active recombinant expressed Saccharomyces cerevisiae transformed cDNA. large-scale fermentation broths using ultrafiltration gel filtration chromatography on Sephacryl S200 HR columns. NH -terminal amino acid sequencing showed cleavage point for activation of pro-enzyme identical F. -produced mature behaved similarly when analysed SDS-PAGE, immunoblotting zymography also cleaved peptides containing Finally, fibrinogen form fibin clot, property we described