Solubilization of serotonin1a and serotonin1b binding sites from bovine brain.
Solubilization of serotonin1a and serotonin1b binding sites from bovine brain.
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牛脑中 5-羟色胺 1a 和 5-羟色胺 1b 结合位点的溶解。
DOI：
10.1111/j.1471-4159.1987.tb05691.x
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发表时间：
1987
影响因子：
4.7
通讯作者：
Shih,JC
中科院分区：
文献类型：
--
作者：
Asarch,KB;Shih,JC
使用0.1% Nonidet P-40和0.3%毛地黄皂苷在含0.1%抗坏血酸的低盐缓冲液中的混合物，从牛脑皮质中溶解5-羟色胺（5-羟色胺1，5-HT 1）结合位点。[3 H]5-HT对可溶性皮质结合位点的亲和力（2.1 nM）与其对膜结合位点的亲和力（2.1 nM）相同。[3H]8- 羟基-2-（二正丙基氨基）四氢喹啉（[3 H]DPAT）是一种选择性5-HT 1a放射性配体，也以与其在粗膜中的亲和力（1.7 nM）相当的高亲和力（1.8 nM）与可溶性皮质结合位点结合。β-羟色胺能药物与[3 H]5-HT结合和[3 H]DPAT与粗品和可溶性膜结合的等级效力存在显著相关性。溶解皮质膜中[3 H]DPAT结合位点相对于[3 H]5-HT位点的密度（35%）与通过螺哌隆置换测定的粗膜中5-HT 1a位点的比例（33%）一致，表明5-HT 1a和5-HT 1b结合位点均已共溶解。[3H]5可溶性制剂中的-HT结合被GTP抑制，表明受体复合物可能已被溶解。在该制剂中未检测到[3 H]螺哌隆特异性结合，表明5-HT 2位点未共溶解。
Serotonin, (5‐hydroxytryptamine1, 5‐HT1) binding sites have been solubilized from bovine brain cortex using a mixture of 0.1 % Nonidet P‐40 and 0.3% digitonin in a low‐salt buffer containing 0.1 % ascorbic acid. The affinity of [3H]5‐HT for the soluble cortical binding sites (2.1 nM) is identical to its affinity at membrane‐bound binding sites (2.1 nM). [3H]8‐Hydroxy‐2‐(di‐n‐propylamino)tetraIin ([3H]DPAT), a selective 5‐HT1a, radioligand, also binds to soluble cortical binding sites with high affinity (1.8 nM) comparable with its affinity in the crude membranes (1.7 nM). A significant correlation exists in the rank order potency of serotonergic agents for [3H]5‐HT binding and for [3H]DPAT binding to crude and soluble membranes. The density of [3H]DPAT binding sites relative to the [3H]5‐HT sites in the solubilized cortical membranes (35%) corresponds well with the proportion of 5‐HT1asites in the crude membranes determined by spiperone displacement (33%), suggesting that both the 5‐HT1aand 5‐HT1bbinding sites have been cosolubilized. [3H]5‐HT binding in the soluble preparations was inhibited by GTP, suggesting that a receptor complex may have been solubilized. [3H]Spiperone‐specific binding was not detectable in this preparation, suggesting that 5‐HT2sites were not cosolubilized.