ENDOPEPTIDASE AND CARBOXYPEPTIDASE ACTIVITIES IN HUMAN URINE WHICH HYDROLYZE BRADYKININ

ENDOPEPTIDASE AND CARBOXYPEPTIDASE ACTIVITIES IN HUMAN URINE WHICH HYDROLYZE BRADYKININ

Author Casarini, Dulce Elena Autor UNIFESP Google Scholar
Alves, Kaethy Bisan Autor UNIFESP Google Scholar
Araujo, M. S. Google Scholar
Stella, Regina Celes de Rosa Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Abstract 1. We have fractionated the bradykinin inactivating activity of human urine by stepwise elution chromatography on DEAE-cellulose and recovered 95% of the inactivating activity and 29% of the protein (absorbance at A280 nm).2. Seven of nine fractions which presented activity were also tested for angiotensin I and II inactivating activity, angiotensin converting activity and for the hydrolysis of hippuryl-His-Leu and hippuryl-Arg. Sites of hydrolysis in bradykinin were determined by HPLC of the hydrolysates and fragments were compared with authentic peptides.3. Cleavage sites demonstrated for Fractions A through G were: Phe8-Arg9 (A and B), Phe5-Ser6 (C and F), Pro7-Phe8 (D), Gly4-Phe5 and Pro7-Phe8 (E) and Pro3-Gly4 (G).4. The relative molecular weight of the bradykininase activity present in each fraction, determined by gel filtration, was: 16 kDa (A), 70 kDa (B), 60 kDa (C), 88 kDa (D), 230 kDa (E), 45 kDa (F) and 49 kDa (G).5. Bradykinin inactivating activity was inhibited 50-100% by 3 mMEDTA (A,B,D,E and G), 1 mMM 2-mercaptoethanol (A, B, C and G), 0.1-mu-M Hg2+ (A, C and G), 0.1 mM PMSF (C and F), 1 mM TPCK (C and F), 1 mM Zn2+ (C), 60-mu-M BPP5a and 40-mu-M BPP9a (D), 0.1-mu-M phosphoramidon (E) and 3 mM sodium p-hydroxymercuribenzoate (G).6. The properties of some of these bradykinin inactivating activities correspond to enzymes previously described in urine and tissues: carboxypeptidases (Fractions A and B), angiotensin I converting enzyme (Fraction D), neutral endopeptidase (Fraction E). However, the chymotrypsin-like activity of Fractions C and F and the prolylendopeptidase activity of Fraction G have not been described before in urine and they are being purified in order to obtain a more accurate characterization.
Keywords HUMAN URINE ENDOPEPTIDASE
CARBOXYPEPTIDASE
KININASE ACTIVITY
BRADYKININ
ANGIOTENSIN CONVERTING ENZYME
BRADYKININ POTENTIATING PEPTIDES
PHENYLMETHANESULFONYL FLUORIDE
PHOSPHORAMIDON
Language English
Date 1992-01-01
Published in Brazilian Journal Of Medical And Biological Research. Sao Paulo: Assoc Bras Divulg Cientifica, v. 25, n. 3, p. 219-229, 1992.
ISSN 0100-879X (Sherpa/Romeo, impact factor)
Publisher Assoc Bras Divulg Cientifica
Extent 219-229
Access rights Closed access
Type Article
Web of Science ID WOS:A1992HN91100001
URI http://repositorio.unifesp.br/11600/43018

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