CYTOSOL ACTIVATOR OF RAT RETICULOCYTE ADENYLATE CYCLASE

这项研究项目的目的是建立身份， 腺苷酸类细胞调节剂的作用机制及其重要性 从大鼠网织红细胞胞浆中获得的环化酶。网织红细胞 胞浆激活蛋白(RCAP)，增强儿茶酚胺敏感性 大鼠网织红细胞膜上的腺苷环化酶只有部分 提纯和鉴定。研究计划的目标是净化 并对RCAP进行了完整的表征；阐明了RCAP的作用机制 它如此深刻地影响儿茶酚胺的反应性；并建立 RCAP在多大程度上和以什么方式调节腺苷酸环化酶 在其他组织和物种中的活性。RCAP将被提纯到 使用成熟的生化技术进行均质，包括制备 等电聚焦和方法中概述的其他具体方法。 RCAP的作用机制将通过以下研究进行探讨 RCAP对荷尔蒙单位的三个组成部分的影响 β-肾上腺素能受体、鸟嘌呤核苷酸偶联蛋白和 腺苷环化酶的催化单位以及它们之间的相互作用 他们。具体方法将包括放射性配基结合研究， 腺苷环化酶活力的测定及鸟嘌呤的评价 通过ADP-核糖化试验检测核苷酸结合蛋白。这个 RCAP对其他刺激或抑制腺苷的激素的影响 环化酶在其他几种哺乳动物和组织中也会 进行调查，以确定RCAP作为一个整体的重要性 荷尔蒙活动的调节器。

The objective of this research project is to establish the identity, mechanism of action, and importance of the cellular regulator of adenylate cyclase obtained from the cytosol of rat reticulocytes. The reticulocyte cytosol activator protein (RCAP), that augments catecholamine-sensitive adenylate cyclase in rat reticulocyte membranes, has been only partially purified and characterized. The goals of the research plan are to purify and to completely characterize RCAP; to elucidate the mechanisms by which it so profoundly influences catecholamine responsiveness; and to establish to what extent, and by what means, RCAP regulates adenylate cyclase activity in other tissues and species. RCAP will be purified to homogeneity using established biochemical techniques including preparative isoelectric focusing and other specific approaches outlined in Methods. The mechanism of action of RCAP will be investigated by studies directed at RCAP's effects upon each of the three components of the hormonal unit, the beta-adrenergic receptor, the guanine nucleotide coupling protein, and the catalytic unit of adenylate cyclase, as well as the interactions among them. Specific methods will include radioligand binding studies, determination of adenylate cyclase activity, and assessment of the guanine nucleotide binding protein by means of the ADP-ribosylation assay. The effects of RCAP upon other hormones that stimulate or inhibit adenylate cyclase and in several other mammalian and tissues will also be investigated in order to determine the importance of RCAP as a general regulator of hormone action.