Linear V1-vascular vasopressin antagonists suitable for radioiodination, biotinylation, and fluorescent labeling.
Linear V1-vascular vasopressin antagonists suitable for radioiodination, biotinylation, and fluorescent labeling.
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线性 V1-血管加压素拮抗剂，适用于放射性碘标记、生物素化和荧光标记。
DOI：
10.1152/ajpendo.1993.265.6.e906
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Madhun,Z
中科院分区：
文献类型：
--
作者：
Thibonnier,M;Bayer,AL;Madhun,Z
我们修改了几个线性V1-血管精氨酸加压素（AVP）拮抗剂，以获得适合于放射性碘标记，生物素化和荧光标记的化合物。在与人血小板V1-血管AVP受体的结合竞争实验中，线性V1拮抗剂苯乙酰基-D-Tyr（Et）-Phe-Gln-Asn-Lys-Pro-Arg-NH 2（PhaaGln）显示出最大的亲和力[解离常数（Kd）= 0.05 +/- 0.01 nM]。放射性碘标记化合物苯乙酰基-D-Tyr（Et）-Phe-Val-Asn-Lys-Pro-125 I标记的Tyr-NH 2（125 I标记的TyrPhaa）的特征在于高亲和力（Kd = 1.42 +/- 0.19 nM）、低非特异性结合和良好的稳定性。PhaaGln偶联十二碳配体保留了对V1血管AVP受体的良好亲和力（Kd = 1.41 +/- 0.20 nM）。复合物PhaaGln-dodecabiotin-抗生物素蛋白是双功能的，因为抗生物素蛋白-琼脂糖柱特异性结合用125 I-TyrPhaa-dodecabiotin标记的V1-血管AVP受体。在A7 r5血管平滑肌细胞加载fura-2，PhaaGln和PhaaGln-dodecabiotin是纯拮抗剂，因为它们阻断AVP诱导的钙动员，但不引起钙信号本身。A7 r5血管平滑肌细胞的V1-血管AVP受体通过结合的PhaaGln-十二碳生物素可视化，所述结合的PhaaGln-十二碳生物素通过用荧光素抗生物素蛋白标记而发荧光。因此，线性V1-血管AVP拮抗剂可用作高亲和力和特异性放射性碘标记、生物素标记和荧光探针，以探索人和动物来源的V1-血管AVP受体。
We modified several linear V1-vascular arginine vasopressin (AVP) antagonists to obtain compounds suitable for radioiodination, biotinylation, and fluorescent labeling. In binding competition experiments with human platelet V1-vascular AVP receptors, the linear V1 antagonist phenylacetyl-D-Tyr(Et)-Phe-Gln-Asn-Lys-Pro-Arg-NH2 (PhaaGln) displayed the greatest affinity [dissociation constant (Kd) = 0.05 +/- 0.01 nM]. The radioiodinated compound phenylacetyl-D-Tyr(Et)-Phe-Val-Asn-Lys-Pro-125I-labeled Tyr-NH2 (125I-labeled TyrPhaa) was characterized by a high affinity (Kd = 1.42 +/- 0.19 nM), a low nonspecific binding, and good stability. PhaaGln coupled to dodecabiotin retained a good affinity for V1-vascular AVP receptors (Kd = 1.41 +/- 0.20 nM). The complex PhaaGln-dodecabiotin-avidin is bifunctional, since an avidin-agarose column specifically bound V1-vascular AVP receptors labeled with 125I-TyrPhaa-dodecabiotin. In A7r5 vascular smooth muscle cells loaded with fura-2, PhaaGln and PhaaGln-dodecabiotin were pure antagonists as they blocked AVP-induced calcium mobilization but did not elicit a calcium signal by themselves. V1-vascular AVP receptors of A7r5 vascular smooth muscle cells were visualized by bound PhaaGln-dodecabiotin made fluorescent by labeling with fluorescein avidin. Thus linear V1-vascular AVP antagonists can be used as high affinity and specificity radioiodinated, biotinylated, and fluorescent probes to explore V1-vascular AVP receptors of human and animal origin.