title: [Structure and function of the receptor for human atrial natriuretic peptide in cultured human skin fibroblasts].
abstract: Cultured human skin fibroblasts possessed the high-affinity and low-capacity binding sites for [125I]alpha-human atrial natriuretic peptide (hANP), in which the dissociation constant and maximal binding capacity were computed to 68.7 +/- 11.3 pM and 7.3 +/- 1.2 fmols/mg protein, respectively, from Scatchard plot analysis. The specific [125I] alpha-hANP binding sites of cultured human fibroblast were displaced by unlabeled atriopeptin I, a truncated analogue, to the same extent as the case of alpha-hANP. In human adrenal membrane fractions, [125I] alpha-hANP binding sites were suppressed only by unlabeled alpha-hANP, while the high concentrations of atriopeptin I could slightly inhibit the binding sites for alpha-hANP. As it was reported that atriopeptin I had more significant affinity to the low-molecular weight ANP receptor (60-70 KD) than that to the high-molecular weight form (130-140 KD), the specific bindings may be attributed by the low-molecular weight ANP receptor in cultured human fibroblasts. Furthermore, alpha-hANP up to 10(-8)M failed to induce the significant cGMP formation in cultured human skin fibroblasts. The molecular weight of [125I]alpha-hANP binding sites of human fibroblasts was identified only at the region of 67 KD and no radioactive band was visualized around the region of large molecular weight ANP receptor in the SDS gel electrophoresis of a crosslinked [125I]alpha-hANP-receptor complex. In contrast, the affinity labeling of [125I]alpha-hANP to the human adrenal membrane fractions showed that 135 KD binding sites were responsible to the human adrenal ANP receptor. In conclusion, cultured human skin fibroblasts have a high-affinity low-capacity receptor for ANP. The molecular weight of ANP receptor is approximately 67 KD, and ANP-specific guanylate cyclase may not be linked to the receptor, suggestive that so-called C receptor may be localized in cultured human fibroblasts.
authors.affiliation: Third Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka.
authors.forename: M
authors.initials: M
authors.lastname: Ohashi
authors.forename: T
authors.initials: T
authors.lastname: Hashiguchi
authors.forename: R
authors.initials: R
authors.lastname: Takayanagi
authors.forename: M
authors.initials: M
authors.lastname: Haji
authors.forename: H
authors.initials: H
authors.lastname: Nawata
mesh_terms: D000311:Adrenal Glands / Q000378:metabolism; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D001665:Binding Sites; D002478:Cells, Cultured; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D008297:Male; D008970:Molecular Weight; D017461:Receptors, Atrial Natriuretic Factor; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000502:physiology*; D012867:Skin / Q000166:cytology / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D011956:Receptors, Cell Surface; D009320:Atrial Natriuretic Factor; D017461:Receptors, Atrial Natriuretic Factor