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A cysteine proteinase in the cercariae ofDiplostomum pseudospathaceum (Trematoda, Diplostomatidae)

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Abstract

A cysteine proteinase was detected in extracts from cercariae of the trematodeDiplostomum pseudospathaceum. The enzyme preferred protein substrates over synthetic, chromogenic peptides. The optimal pH for hydrolysis of substrates was 7.2 for azocoll, 6.4 and 7.6 for azocasein, 7.6 for azoalbumin, and 6.8 forN-benzoyl-l-arginine-4-nitroanilide. Elastin-Congo red and certainN-blockedl-aminoacyl-andl-peptidyl nitroanilides bearingl-phenylalanine,l-alanine,l-tyrosine, andl-leucine at the P1 subsite were not hydrolyzed. Thiol-reducing and divalent cation-complexing agents stimulated the proteinase activity, whereas thiol-blocking agents inhibited it. The relative molecular weight of the enzyme was approximately 40 000 as determined by SDS-PAGE. Detection of an identical proteinase in water after treatment of living cercariae with praziquantel suggests that the enzyme occupied the penetration glands in the larvae. Thus, when secreted by the parasite during invasion of an appropriate host, the enzyme might act as a penetration-promoting factor.

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Abbreviations

SDS-PAGE :

sodium dodecyl sulfatepolyacrylamide electrophoresis

BSA :

bovine serum albumin

TRIS :

tris(hydroxymethyl)aminomethane

EGTA :

ethylene glycol-O,OO′-bis(2-aminoethyl)N,N,N′,N′-tetraacetic acid

DTE :

dithioerythritol

2-ME :

2-mercaptoethanol

pHMB :

sodiump-hydroxymercuribenzoate

PMSF :

phenylmethanesulfonyl fluoride

TPCK N :

tosyl-l-phenylalanine chloromethyl ketone

LCK Nα :

tosyl-l-lysine chloromethyl ketone

SBTI :

soybean trypsin inhibitor

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Authors and Affiliations

  1. Institute of Parasitology, Polish Academy of Sciences, Pasteura 3, P.O.B. 153, PL-00-973, Warsaw, Poland

    T. Moczoń

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  1. T. Moczoń
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This work was supported by research grant PB 6 6004 92 03 from the State Committee for Scientific Research

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Moczoń, T. A cysteine proteinase in the cercariae ofDiplostomum pseudospathaceum (Trematoda, Diplostomatidae) . Parasitol Res 80, 680–683 (1994). https://doi.org/10.1007/BF00932952

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  • DOI: https://doi.org/10.1007/BF00932952

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