title: Liver bradykinin-inactivating-endopeptidase is similar to the metalloendopeptidase (EC 3.4.24.15).
abstract: The bradykinin-inactivating-endopeptidase (BIE) removal from rat liver, by perfusing the organ with 0.05% Triton X-100, achieved its maximum at 10 min of perfusion and falls to 50% of the maximum in 30 min, a pattern similar to AST removal. Using an internally quenched fluorescent BK analogue (Abz-RPPGFSPFRQ-EDDnp) we further characterized this enzyme: it is activated by low concentrations of 2-mercaptoethanol, inhibited by p-hydroxymercuribenzoate, o-phenanthroline and EDTA, and is resistant to enalapril, E-64 and PMSF. These results suggest that BIE is a metalloendopeptidase containing a thiol group important for its activity. BIE also hydrolyses the peptides Abz-GGFLRRVQ-EDDnp, Abz-GPQGLAGQ-EDDnp, Abz-FRSVQ-EDDnp, and Abz-ARVRRANSFLQ-EDDnp. All these properties are very similar to those described or assayed by us for EC 3.4.24.15, isolated initially from rat testes and then from several organs of different animals. Both BIE and EC 3.4.24.15: hydrolyze the F5S6 bond of the BK fluorescent substrate; are efficiently inhibited by Orlowski specific inhibitor (CFP-AAF-pAB, Ki 4.4 x 10(-7) M and 1.25 x 10(-7) M, respectively); have the same electrophoretic mobility in SDS-PAGE (Mr 78,000); and are both recognized by three polyclonal antibodies raised against rat testes EC 3.4.24.15. In conclusion, BIE appears to be EC 3.4.24.15.
authors.affiliation: Department of Biochemistry, Universidade Federal de São Paulo/EPM, Brazil.
authors.forename: H M
authors.initials: HM
authors.lastname: Molina
authors.forename: A K
authors.initials: AK
authors.lastname: Carmona
authors.forename: M
authors.initials: M
authors.lastname: Kouyoumdjian
authors.forename: D R
authors.initials: DR
authors.lastname: Borges
authors.forename: L
authors.initials: L
authors.lastname: Juliano
mesh_terms: D000818:Animals; D010450:Endopeptidases / Q000032:analysis* / Q000502:physiology; D008099:Liver / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000032:analysis* / Q000502:physiology; D051381:Rats; D013379:Substrate Specificity
chemical_list: D010450:Endopeptidases; D008666:Metalloendopeptidases; C032615:bradykinin kininase