Kininogenase activity by the major cysteinyl proteinase (Cruzipain) from Trypanosoma cruzi

Kininogenase activity by the major cysteinyl proteinase (Cruzipain) from Trypanosoma cruzi

Author Del Nery, Elaine Autor UNIFESP Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Lima, Ana Paula CA Google Scholar
Scharfstein, Julio Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Universidade Federal do Rio de Janeiro (UFRJ)
Abstract The major isoform of Trypanosona cruzi cysteinyl proteinase (cruzipain) has generated Lys-bradykinin (Lys-BK or kallidin), a proinflammatory peptide, by proteolysis of kininogen. the releasing of this peptide was demonstrated by mass spectrometry, radioimmunoassay, and ileum contractile responses, the kinin-releasing activity was immunoabsorbed selectively by monoclonal antibodies to the characteristic COOH-terminal domain of cruzipain. To determine the hydrolysis steps that account for the kininogenase activity of cruzipain, we synthesized a fluorogenic peptide (o-aminobenzoyl-Leu-Gly-Met-Ile-Ser-Leu-Met-Lys-Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg(389)-Ser(390)-Ser-Arg-Ile-NH2) based on the sequence Leu(373) to Ile(393) of the human high molecular weight kininogen, the hydrolysis products from this peptide were isolated by high performance liquid chromatography, and Lys-BK was characterized as the major released kinin by mass spectrometry. Intramolecularly quenched fluorogenic peptides spanning the Met(379)-Lys(380) and Arg(389)-Ser(390) bradykinin-flanking sequences were then used to assess the substrate specificity re requirements of the parasite-derived protease compared with two COOH-terminal truncated recombinant isoforms (cruzain and cruzipain 2). in contrast to the high catalytic efficiency of parasite-derived cruzipain, the recombinant proteinases cleaved the bradykinin-flanking sites at markedly different rates. in addition, we also demonstrated that cruzipain activates plasmatic prekallikrein, which would be a second and indirect way of the parasite protease to release bradykinin.
Language English
Date 1997-10-10
Published in Journal of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 272, n. 41, p. 25713-25718, 1997.
ISSN 0021-9258 (Sherpa/Romeo, impact factor)
Publisher Amer Soc Biochemistry Molecular Biology Inc
Extent 25713-25718
Access rights Open access Open Access
Type Article
Web of Science ID WOS:A1997YA35800051

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