Comparison of binding and cyclic GMP accumulation by atrial natriuretic peptides in endothelial cells.
Comparison of binding and cyclic GMP accumulation by atrial natriuretic peptides in endothelial cells.
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DOI：
10.1016/0167-4889(86)90040-6
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发表时间：
1986-01
期刊：
影响因子：
--
通讯作者：
Dale C. Leitmans;F. Murad
中科院分区：
文献类型：
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作者：
Dale C. Leitmans;F. Murad
Rat125I 标记的心房钠尿因子 (ANF (8–33)) 用于鉴定培养的牛主动脉内皮细胞上的 ANF 受体。 125 I-ANF在37℃下与汇合内皮细胞的特异性结合是可饱和的并且具有高亲和力。平衡结合数据的 Scatchard 分析表明，内皮细胞含有 aKdof 0.1 ± 0.01 nM 的一类结合位点。这种特殊的内皮细胞克隆每个细胞有 16 000 ± 1300 个受体。与 125 I-ANF 结合竞争的效力顺序为人心房钠尿肽 (hANP)= 心房钠尿因子 (ANF (8-33))> 心房肽 II > 心房肽 III > 心房肽。最弱的竞争对手心肽素 I 的 KI 为 0.45 nM，仅比 hANP 和 ANF 的 KI 高 6 倍 (8-33)。 ANF (8-33) 和 hANP 在 0.5 mM 异丁基甲基黄嘌呤存在下，在 10 pM 时使环 GMP 含量增加 15-20 倍，在 10 nM 时使环 GMP 含量最大增加 500 倍。引起 hANP、ANF (8-33)、atriopeptin I、atriopeptin II 和 atriopeptin III 环 GMP 半最大增加所需的浓度分别为 0.30、0.35、>500、4.0 和 5.0 nM。尽管 atriopeptin I 起到部分激动剂的作用，但它无法拮抗 ANF (8-33) 对环 GMP 形成的影响。这些发现表明内皮细胞具有多个且功能不同的 ANF 结合位点。
Rat125I-labeled atrial natriuretic factor (ANF (8–33)) was used to identify ANF receptors on cultured bovine aortic endothelial cells. Specific binding of125I-ANF at 37°C to confluent endothelial cells was saturable and of high affinity. Scatchard analysis of the equilibrium binding data indicated that endothelial cells contain a single class of binding sites with aKdof 0.1 ± 0.01 nM. This particular clone of endothelial cells had 16 000 ± 1300 receptors per cell. The order of potency for competing with125I-ANF binding was human atrial natriuretic peptide (hANP)= atrial natriuretic factor (ANF (8–33))>atriopeptin II > atriopeptin III > atriopeptin. The weakest competitor, atriopeptin I, had aKIof 0.45 nM, which was only 6-fold higher than theKIfor hANP and ANF (8–33). ANF (8–33) and hANP in the presence of 0.5 mM isobutylmethylxanthine produced a 15–20-fold increase in cyclic GMP content at 10 pM and a maximal 500-fold elevation of cyclic GMP at 10 nM. The concentrations required to elicit a half-maximal increase in cyclic GMP for hANP, ANF (8–33), atriopeptin I, atriopeptin II and atriopeptin III were 0.30, 0.35, >500, 4.0 and 5.0 nM, respectively. Although atriopeptin I acted as a partial agonist, it was unable to antagonize the effect of ANF (8–33) on cyclic GMP formation. These findings suggest that endothelial cells have multiple and functionally distinct ANF-binding sites.