title: Highly selective hydrolysis of kinins by recombinant prolylcarboxypeptidase.
abstract: We have previously cloned a cDNA encoding human prolylcarboxypeptidase (PRCP) and expressed the cDNA in the Schneider 2 (S2) drosophila cell line. Here, we further characterized this recombinant enzyme. Investigations were performed to determine whether recombinant PRCP (rPRCP) metabolizes kinins (BK 1-9 and BK 1-8). The metabolites of these kinins were identified by LC/MS. rPRCP metabolized BK 1-8 to BK 1-7, whereas rPRCP was ineffective in metabolizing BK 1-9. The hydrolysis of BK 1-8 by rPRCP was dose- and time-dependent. A homology model of PRCP was developed based upon the sequence of dipeptidyl-peptidase 7 (DPP7, PDB ID: 3JYH), and providentially, the structure of PRCP (PDB ID: 3N2Z) was characterized during the course of our investigation. Docking studies of bradykinin oligopeptides were performed both from the homology model, and from the crystal structure of PRCP. These docking studies may provide a better understanding of the contribution of specific residues involved in substrate selectivity of human PRCP.
authors.affiliation: Department of Medicinal Chemistry, School of Pharmacy, University of Mississippi, University, MS 38677-1848, USA.
authors.forename: S M
authors.initials: SM
authors.lastname: Chajkowski
authors.forename: J
authors.initials: J
authors.lastname: Mallela
authors.forename: D E
authors.initials: DE
authors.lastname: Watson
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: C R
authors.initials: CR
authors.lastname: McCurdy
authors.forename: J M
authors.initials: JM
authors.lastname: Rimoldi
authors.forename: Z
authors.initials: Z
authors.lastname: Shariat-Madar
mesh_terms: D000818:Animals; D001920:Bradykinin / Q000096:biosynthesis / Q000737:chemistry; D002268:Carboxypeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020134:Catalytic Domain; D002853:Chromatography, Liquid; D018076:DNA, Complementary / Q000235:genetics; D004330:Drosophila; D006801:Humans; D006860:Hydrogen Bonding; D006868:Hydrolysis; D007705:Kinins / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D007705:Kinins; D011994:Recombinant Proteins; D002268:Carboxypeptidases; C022448:lysosomal Pro-X carboxypeptidase; D001920:Bradykinin