title: Role of "handle" region of prorenin prosegment in the non-proteolytic activation of prorenin by binding to membrane anchored (pro)renin receptor.
abstract: A role of the "handle" region in the prorenin prosegment sequence was investigated to demonstrate the crucial non-proteolytic activation of prorenin by binding to the recombinant (pro)renin receptor on the COS-7 cell membrane. The plasmid DNA containing either rat or human (pro)renin receptor was transfected into the COS-7 cells. The highest amount of receptor was observed on the COS-7 cell membrane after 18 h transfection. Of the total rat and human prorenin, 90% and 50% were bound to each of the respective receptors, respectively. The Kd values were 0.89 and 1.8 nM, respectively. Rat prorenin was activated non-proteolytically by the receptor. The Km was determined 1.0 microM when sheep angiotensinogen was used as the substrate. Human prorenin was also activated by the receptor. The Km was 0.71 microM. Additionally, decapeptides (10P-19P) known as "decoy" peptide and pentapeptides (11P-15P) named "handle" region peptide, were observed to inhibit the binding of both prorenins to receptors, respectively. The Ki were similar around 7 nM for both the peptides. Other two region peptides in the prosegment did not interfere the binding. These results show that the "handle" region probably plays a crucial role in prorenin binding to the receptor and in its enzymic activity by non-proteolytic activation.
authors.affiliation: Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
authors.forename: Nabi A H M
authors.initials: NA
authors.lastname: Nurun
authors.forename: Nasir M
authors.initials: NM
authors.lastname: Uddin
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Iwata
authors.forename: Atsuhiro
authors.initials: A
authors.lastname: Ichihara
authors.forename: Tadashi
authors.initials: T
authors.lastname: Inagami
authors.forename: Fumiaki
authors.initials: F
authors.lastname: Suzuki
mesh_terms: D020816:Amino Acid Motifs; D000808:Angiotensinogen / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002522:Chlorocebus aethiops; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D017434:Protein Structure, Tertiary; D051381:Rats; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D012083:Renin / Q000737:chemistry* / Q000378:metabolism; D012756:Sheep; D000093342:Prorenin Receptor
chemical_list: D011956:Receptors, Cell Surface; D000808:Angiotensinogen; D012083:Renin; D000093342:Prorenin Receptor