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A cysteine proteinase in the penetration glands of the cercariae of Cotylurus cornutus (Trematoda, Strigeidae).

Moczoń T - Parasitol. Res. (2010)

Bottom Line: 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 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.

View Article: PubMed Central - PubMed

Affiliation: W Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818 Warszawa, Poland. moczon@twarda.pan.pl

ABSTRACT
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.

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Hydrolysis of Z-Arg-NHnapMeO by the cysteine proteinase occupying the penetration glands (pg) and the secretory canals (sc) of the cercariae examined with the fluorescence method vs. ventral sucker; t, tail
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Fig1: Hydrolysis of Z-Arg-NHnapMeO by the cysteine proteinase occupying the penetration glands (pg) and the secretory canals (sc) of the cercariae examined with the fluorescence method vs. ventral sucker; t, tail

Mentions: Golden-yellow fluorescing precipitates were observed in those areas of the body of the cercariae which were occupied by the penetration glands and by the secretory canals, both being filled with the secretion produced by the glandular cells (Fig. 1). The intensity of the reaction increased with the elevation of the pH, becoming optimal at the pH 5.6, the highest value at which the reaction product remains microcrystalline for a number of minutes and satisfactorily reflects the examined structures; however, from an unknown reason only one in approximately 100 of the examined specimens revealed a satisfactory pattern of the reaction. Due to the fixation and defatting, as well as to a low magnifying power, the activity of cathepsin B in the whole larval body was poorly perceptible.Fig. 1


A cysteine proteinase in the penetration glands of the cercariae of Cotylurus cornutus (Trematoda, Strigeidae).

Moczoń T - Parasitol. Res. (2010)

Hydrolysis of Z-Arg-NHnapMeO by the cysteine proteinase occupying the penetration glands (pg) and the secretory canals (sc) of the cercariae examined with the fluorescence method vs. ventral sucker; t, tail
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3040816&req=5

Fig1: Hydrolysis of Z-Arg-NHnapMeO by the cysteine proteinase occupying the penetration glands (pg) and the secretory canals (sc) of the cercariae examined with the fluorescence method vs. ventral sucker; t, tail
Mentions: Golden-yellow fluorescing precipitates were observed in those areas of the body of the cercariae which were occupied by the penetration glands and by the secretory canals, both being filled with the secretion produced by the glandular cells (Fig. 1). The intensity of the reaction increased with the elevation of the pH, becoming optimal at the pH 5.6, the highest value at which the reaction product remains microcrystalline for a number of minutes and satisfactorily reflects the examined structures; however, from an unknown reason only one in approximately 100 of the examined specimens revealed a satisfactory pattern of the reaction. Due to the fixation and defatting, as well as to a low magnifying power, the activity of cathepsin B in the whole larval body was poorly perceptible.Fig. 1

Bottom Line: 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 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.

View Article: PubMed Central - PubMed

Affiliation: W Stefański Institute of Parasitology, Polish Academy of Sciences, Twarda 51/55, 00-818 Warszawa, Poland. moczon@twarda.pan.pl

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus