title: Substrate specificity of prorenin converting enzyme of mouse submandibular gland. Analysis using site-directed mutagenesis.
abstract: Renin is produced from a larger, inactive precursor, prorenin, through endoproteolytic cleavage at paired basic amino acids. Recently, we have purified and characterized an enzyme, which catalyzes the endoproteolytic process, from mouse submandibular gland. The enzyme, named prorenin converting enzyme, specifically cleaves the peptide bond on the COOH-side of the Arg residue at the Lys-Arg pair of mouse Ren 2 prorenin, but does not cleave mouse Ren 1 and human prorenins. In this study, by synthesizing a series of mutant mouse prorenins using site-directed mutagenesis and the Xenopus oocyte expression system, we have investigated the role of the basic pair as the recognition signal for the enzyme as well as the determinant of the substrate specificity. The results indicate that the basic amino acid at the COOH-side but not at the NH2-side of the basic pair of Ren 2 prorenin is essential for processing directed by prorenin converting enzyme, and that the Arg residue at the COOH-side is more preferable for processing than the Lys. The results also demonstrated that the presence of a Pro residue next to the Lys-Arg pair prevents the processing of Ren 1 prorenin.
authors: Institute of Biological Sciences, University of Tsukuba, Ibaraki, Japan.
authors: K
authors: K
authors: Nakayama
authors: W S
authors: WS
authors: Kim
authors: T
authors: T
authors: Nakagawa
authors: M
authors: M
authors: Nagahama
authors: K
authors: K
authors: Murakami
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002678:Chimera; D003001:Cloning, Molecular; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D013363:Submandibular Gland / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D012333:RNA, Messenger; D012697:Serine Endopeptidases; C063570:prorenin Ren2 converting enzyme