title: The His-Pro-Phe motif of angiotensinogen is a crucial determinant of the substrate specificity of renin.
abstract: The amino acid sequence His-Pro-Phe as N-terminal residues 6-8 of the natural renin substrate, angiotensinogen, is conserved among species. We investigated whether this His-Pro-Phe motif functions as the determinant of the substrate specificity of renin. Mutant angiotensinogens in which the Ile-His-Pro-Phe-His-Leu sequence at positions 5-10 of wild-type angiotensinogen was replaced by either His-Pro-Phe-His-Leu-Leu or Ala-Ile-His-Pro-Phe-His were cleaved by renin at the C-terminal side of residues 9 and 11, respectively, while wild-type angiotensinogen was cleaved at residue 10. A triple Ala substitution for the His-Pro-Phe motif of angiotensinogen prevented its cleavage by renin. In contrast, triple Ala substitution for residues 9-11, including the natural site of cleavage by renin, allowed cleavage between the two Ala residues at positions 10 and 11. Furthermore, the 33-residue C-terminal peptide of human megsin, which carries a naturally occurring His-Pro-Phe sequence, was cleaved by renin at the C-terminal side of the His-Pro-Phe-Leu-Phe sequence. These results indicate that the His-Pro-Phe motif of angiotensinogen is a crucial determinant of the substrate specificity of renin. By binding to a corresponding pocket on renin, the His-Pro-Phe motif may act as a molecular anchor to recruit the scissile peptide bond to a favorable site for catalysis.
authors.affiliation: Laboratory of Applied Biochemistry, Faculty of Applied Biological Science, Gifu University, 1-1 Yanagido, Gifu 501-1193, Japan. nakagawa@gifu-u.ac.jp
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Jyunji
authors.initials: J
authors.lastname: Akaki
authors.forename: Ryousuke
authors.initials: R
authors.lastname: Satou
authors.forename: Masatoshi
authors.initials: M
authors.lastname: Takaya
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Iwata
authors.forename: Akemi
authors.initials: A
authors.lastname: Katsurada
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Nishiuchi
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Ohmura
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Suzuki
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakamura
mesh_terms: D000595:Amino Acid Sequence; D000808:Angiotensinogen / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D001001:Anura; D002417:Cattle; D017124:Conserved Sequence; D004285:Dogs; D005399:Fishes; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012083:Renin / Q000737:chemistry*; D016415:Sequence Alignment; D012756:Sheep; D013379:Substrate Specificity; D013879:Thioredoxins / Q000737:chemistry; D013997:Time Factors
chemical_list: D011993:Recombinant Fusion Proteins; D000808:Angiotensinogen; D013879:Thioredoxins; D012083:Renin