Detection and preliminary characterization of Taenia solium metacestode proteases

A. Clinton White, J. L. Molinari, A. V. Pillai, A. A. Rege

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

The metacestode of Taenia solium persists for years in the human central nervous system. As proteolytic enzymes play an important role in the survival of tissue helminths, we examined extracts of T. solium metacestodes for proteolytic activity using 9 synthetic peptide substrates and 3 proteins (hemoglobin, albumin, and immunoglobulin G). The proteolytic enzymes were classified based on their inhibitor profiles. At neutral pH, aminopeptidase (arginine-7-amino-4-trifluoromethylcoumarin) and endopeptidase (benzyloxy- carbonyl-glycine-glycine-arginine-7-amino-4-trifluoromethylcoumarin) substrates were cleaved. Hydrolysis of both substrates was inhibited by chelating agents, which inhibit metalloproteases. Peak activity with both substrates eluted in gel filtration fractions corresponding to a molecular weight of about 104 kDa. Cysteine protease activity was identified, which cleaved benzyloxy-carbonyl-phenylalanine-arginine-7-amino-4- trifluoromethylcoumarin (Z-Phe-Arg-AFC) and hemoglobin. Cleavage of Z-Phe- Arg-AFC was maximal at acid pH, was stimulated by thiols, and was inhibited by leupeptin and Ep459. Peak cysteine protease activity eluted in gel filtration fractions corresponding to a molecular weight of 32 kDa. Aspartic protease activity was identified by specific inhibition with pepstatin of acid digestion of hemoglobin and immunoglobulin G. Immunoglobulin digestion occurred at acid pH, with preferential degradation of the heavy chain. Upon gel filtration chromatography, the aspartic protease activity eluted as a broad peak with maximal activity at about 90 kDa. No serine protease activity was detected. None of the parasite enzymes digested albumin. Proteolytic enzymes of T. solium may be important for parasite survival in the intermediate host, by providing nutrients and digesting host immune molecules.

Original languageEnglish (US)
Pages (from-to)281-287
Number of pages7
JournalJournal of Parasitology
Volume78
Issue number2
StatePublished - 1992
Externally publishedYes

Fingerprint

Taenia solium
metacestodes
Peptide Hydrolases
hemoglobin
proteinases
enzyme
arginine
aspartic proteinases
substrate
gel
cysteine proteinases
immunoglobulin G
Gel Chromatography
enzymes
glycine (amino acid)
Hemoglobins
Cysteine Proteases
albumins
acids
parasite

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Microbiology
  • Parasitology

Cite this

Detection and preliminary characterization of Taenia solium metacestode proteases. / White, A. Clinton; Molinari, J. L.; Pillai, A. V.; Rege, A. A.

In: Journal of Parasitology, Vol. 78, No. 2, 1992, p. 281-287.

Research output: Contribution to journalArticle

White, AC, Molinari, JL, Pillai, AV & Rege, AA 1992, 'Detection and preliminary characterization of Taenia solium metacestode proteases', Journal of Parasitology, vol. 78, no. 2, pp. 281-287.
White, A. Clinton ; Molinari, J. L. ; Pillai, A. V. ; Rege, A. A. / Detection and preliminary characterization of Taenia solium metacestode proteases. In: Journal of Parasitology. 1992 ; Vol. 78, No. 2. pp. 281-287.
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