title: Characterization of prorenin activation using a synthetic peptide substrate.
abstract: Human renin is synthesized as an inactive zymogen (prorenin) which is processed to the active form. We synthesized an 11-amino acid peptide which spans the human prorenin processing site in order to develop a simple assay to study human prorenin activation. Six enzymes which are capable of activating recombinant prorenin in vitro were studied. Four of these enzymes digested the synthetic peptide in a specific fashion, as analyzed by reverse-phase high-performance liquid chromatography. Amino acid analysis of the purified digestion products revealed that trypsin cleaves between Arg-Leu, the authentic processing site, while kallikrein, plasmin and elastase all cleaved at alternate sites. On the other hand, pepsin and cathepsin D did not cleave this substrate, suggesting that the activation of prorenin by these proteases might occur at a site distinct from the authentic processing site. Our data suggest that this synthetic peptide may be used as a simple and specific assay for prorenin activation.
authors.affiliation: Falk Cardiovascular Research Center, Stanford, CA 94305-5246.
authors.forename: D
authors.initials: D
authors.lastname: Dubin
authors.forename: R E
authors.initials: RE
authors.lastname: Pratt
authors.forename: K Y
authors.initials: KY
authors.lastname: Hui
authors.forename: V J
authors.initials: VJ
authors.lastname: Dzau
mesh_terms: D000595:Amino Acid Sequence; D002402:Cathepsin D / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D007610:Kallikreins / Q000378:metabolism; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000378:metabolism; D010434:Pepsin A / Q000378:metabolism; D011498:Protein Precursors / Q000138:chemical synthesis*; D012083:Renin / Q000138:chemical synthesis* / Q000378:metabolism*; D013379:Substrate Specificity; D014357:Trypsin / Q000378:metabolism
chemical_list: D004792:Enzyme Precursors; D011498:Protein Precursors; C070714:peptide 8A; D007610:Kallikreins; D010196:Pancreatic Elastase; D014357:Trypsin; D005341:Fibrinolysin; D010434:Pepsin A; D012083:Renin; D002402:Cathepsin D