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人血小板中[3H]丙咪嗪结合位点的增溶及测定

Solubilization and assay of [3H]imipramine binding sites from human platelets.

作者信息

Davis A, Morris J, Tang S W

出版信息

Eur J Pharmacol. 1983 Jan 21;86(3-4):353-9. doi: 10.1016/0014-2999(83)90184-x.

Abstract

[3H]Imipramine binding sites were characterized on outdated (72-96 h) human platelets. The binding site profile was very similar to that previously described for fresh platelets and for CNS membrane preparations. [3H]Imipramine binding sites are believed to represent the recognition site for the 5-hydroxytryptamine uptake system. In order to further investigate this molecular relationship, we solubilized [3H]imipramine binding sites using 1% digitonin and characterised them using a polyethylene glycol precipitation assay. Solubilization did not alter the pharmacological specificity or affinity of the [3H]imipramine binding site and recovery of sites was very high. It was concluded that the solubilization procedure left the binding site remarkably intact. The ready availability of platelets and the convenience of the binding assay makes this system amenable for further purification steps.

摘要

[3H]丙咪嗪结合位点在过期(72 - 96小时)的人血小板上进行了表征。结合位点图谱与先前描述的新鲜血小板和中枢神经系统膜制剂的图谱非常相似。[3H]丙咪嗪结合位点被认为代表5 - 羟色胺摄取系统的识别位点。为了进一步研究这种分子关系,我们用1%的洋地黄皂苷溶解[3H]丙咪嗪结合位点,并使用聚乙二醇沉淀试验对其进行表征。溶解并未改变[3H]丙咪嗪结合位点的药理学特异性或亲和力,且位点回收率非常高。得出的结论是,溶解过程使结合位点保持相当完整。血小板易于获取以及结合测定的便利性使得该系统适合进一步的纯化步骤。

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