title: Qualitative and quantitative analyses of (pro)renin receptor in the medium of cultured human umbilical vein endothelial cells.
abstract: A 30-kDa protein in the medium of cultured human umbilical vein endothelial cells (HUVECs) was identified as (pro)renin receptor, (P)RR, by western blot analysis using anti-human (P)RR antibodies. The protein bound recombinant human prorenin with a K(D) of 4.0 nmol l(-1) and activated prorenin. These observations suggest the presence of soluble (P)RR, s(P)RR, in the medium of cultured HUVECs. For quantification of the s(P)RR in the medium, an enzyme-linked immunosorbent assay (ELISA) was established. The quantitative range of the ELISA was validated over a nominal range of 7.5-300 pmol l(-1) in the wells of a microtiter plate. The assay system showed good linearity (r(2)=0.99) with interassay (5.8-9.7%) and intraassay (2.1-7.0%) precision. Using this method, the concentration of s(P)RR in the culture medium of HUVECs was measured to be 32 pmol l(-1). Therefore, these results show qualitative and quantitative evidence that prorenin can be activated after binding to s(P)RR secreted from cultured HUVECs.
authors.affiliation: Laboratory of Animal Biochemistry, United Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
authors.forename: Kazal Boron
authors.initials: KB
authors.lastname: Biswas
authors.forename: Ahm Nurun
authors.initials: AN
authors.lastname: Nabi
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Arai
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Nakagawa
authors.forename: Akio
authors.initials: A
authors.lastname: Ebihara
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: D002478:Cells, Cultured; D003470:Culture Media; D042783:Endothelial Cells / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D012083:Renin / Q000378:metabolism; D014471:Umbilical Veins / Q000166:cytology / Q000378:metabolism; D000093342:Prorenin Receptor
chemical_list: D003470:Culture Media; D011956:Receptors, Cell Surface; D012083:Renin; D000093342:Prorenin Receptor