A cysteine proteinase in the penetration glands of the cercariae of Cotylurus cornutus (Trematoda, Strigeidae).
A cysteine proteinase from the penetration glands of Cotylurus cornutus cercariae was examined with histochemical and biochemical methods. The enzyme hydrolyzed gelatin, azocoll, azocasein, azoalbumin, N-blocked-L-arginine-4-methoxy-2-naphthylamide, and N-blocked-p-nitroanilide, but did not degrade elastin. The metal ion complexane ethylenediamine tetraacetate and the thiol-reducing compound dithioerythritol enhanced the proteinase activity, whereas the thiol-blocking compounds p-hydroxymercuribenzoate and N-ethylmaleimide (NEM) inhibited it. The enzyme was also sensitive to leupeptin but insensitive to soybean trypsin inhibitor. An electrophoretic separation of extract proteins from the cercariae under acidic, non-denaturing conditions and in the presence of 0.1% gelatin in a polyacrylamide gel revealed the presence of two distinct and three weak transparent bands in the gel resulting from a gelatinolytic activity at pH 6.8. The distinct bands apparently resulted from the activity of the glandular enzyme and lysosomal cathepsin B, whereas the weak ones presumably indicated these enzymes partially degraded in the course of the preparative procedure. No gelatinolysis occurred following treatment of an extract sample with 0.1 mM NEM.
Moczoń
Tadeusz
T
W Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818 Warszawa, Poland. moczon@twarda.pan.pl
D000818:Animals; D002401:Cathepsin B / Q000737:chemistry / Q000378:metabolism; D058487:Cercaria / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D057056:Cysteine Proteases / Q000737:chemistry / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004226:Dithioerythritol / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005033:Ethylmaleimide / Q000494:pharmacology; D006651:Histocytochemistry; D006863:Hydrogen-Ion Concentration; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D014200:Trematoda / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism
D015853:Cysteine Proteinase Inhibitors; D006902:Hydroxymercuribenzoates; D007976:Leupeptins; C030601:4-hydroxymercuribenzoate; D004226:Dithioerythritol; D004492:Edetic Acid; D057056:Cysteine Proteases; D002401:Cathepsin B; C032854:leupeptin; D005033:Ethylmaleimide