CYTOSOL ACTIVATOR OF RAT RETICULOCYTE ADENYLATE CYCLASE

本研究项目的目标是建立身份， 作用机制以及腺苷酸细胞调节剂的重要性 从大鼠网织红细胞的胞浆中获得的环化酶。 网织红细胞 胞质溶胶激活蛋白（RCAP），增强儿茶酚胺敏感性 腺苷酸环化酶在大鼠网织红细胞膜，一直只是部分 纯化和表征。 研究计划的目标是 并完全表征RCAP;阐明其机制， 它如此深刻地影响了儿茶酚胺的反应;并建立 在何种程度上，通过何种方式，RCAP调节腺苷酸环化酶 在其他组织和物种中的活性。 RCAP将被纯化， 使用已建立的生物化学技术，包括制备 等电聚焦和方法中概述的其他特定方法。 RCAP的作用机制将通过以下研究进行研究： RCAP对激素单位的三个组成部分中的每一个的影响， β-肾上腺素能受体，鸟嘌呤核苷酸偶联蛋白，和 腺苷酸环化酶的催化单位，以及 他们 具体方法包括放射性配体结合研究， 腺苷酸环化酶活性测定和鸟嘌呤 通过ADP-核糖基化测定的核苷酸结合蛋白。 的 RCAP对其他刺激或抑制腺苷酸的激素的影响 环化酶和其他几种哺乳动物和组织中也将被 调查，以确定RCAP作为一般的重要性， 激素作用的调节器。

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.