REGULATION OF ADENYLATE CYCLASE

腺苷酸环化酶的调节

• 批准号: 3238351
• 负责人: Roger A Johnson
• 金额: $ 12.92万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3238351
• 关键词: adenylate cyclase; calcium metabolism; cell growth regulation; chromatography; crosslink; cyclic AMP; guanine nucleotides; guanosine triphosphate; hormone regulation /control mechanism; hydrolysis; monoclonal antibody

Adenylate cyclase is the mediator of the action of numerous hormones and neurotransmitters and it is central to the control of metabolism and function in probably all tissues. Its activity is modulated by numerous agents of physiological and biochemical interest, including calcium and some bacterial toxins, and consequently a better understanding of the control of cellular cAMP metabolism is dependent on an understanding of the regulation of adenylate cyclase. As with numerous other enzymes important to cell regulation, calcium activation of a brain adenylate cyclase is mediated by calmodulin, though the basis of the sensitivity of the brain cyclase to calmodulin is not known. There is only an emerging picture of the number of components of the hormone receptor-adenylate cyclase system, without any clear understanding of the mechanisms of their interaction which regulate cyclase activity. The proposed research focuses on the mechanisms of calmodulin-induced activation of adenylate cyclase and will ascertain whether it: a) involves the participation of the stimulatory and inhibitory guanine nucleotide regulatory components; b) involves other unidentified components, unique to the calmodulin-sensitive enzyme; c) resides in unique properties of the catalytic unit of the calmodulin-sensitive enzyme; or d) involves a combination of these possibilities. The two principal approaches will be to purify the Ca2+/calmodulin-sensitive adenylate cyclase and to identify with labelled calmodulin and crosslinking agents calmodulin binding proteins associated with this enzyme. The potential role of Ns and/or Ni on calmodulin activation of the cyclase will be determined through indirect and direct assessments and through reconstitution studies. Given that divalent cation has an established role in the regulation of adenylate cyclase activity, character of their involvement in calmodulin-activation of the brain enzyme will be determined. Polyclonal and/or monoclonal antibodies will be prepared against the cyclase catalytic unit and/or the associated calmodulin-binding protein. These will be used: a) to develop immunopurification adsorbants; and b)\to clarify questions of localization of this form of cyclase or of the associated calmodulin-binding protein. In the narrow sense, the studies will aid our understanding of how individual agents change adenylate cyclase activity, but in the broader sense will give insights into the integration of cell regulatory processes involving both cAMP and calcium.

腺苷酸环化酶是多种激素和 神经递质，这对于控制新陈代谢和 可能在所有组织中起作用。 它的活性由许多人调节 生理和生化兴趣的药物，包括钙和 一些细菌毒素，因此对 蜂窝营地代谢的控制取决于对 腺苷酸环化酶的调节。 与许多其他酶一样重要 对于细胞调节，脑腺苷酸环化酶的钙激活为 通过钙调蛋白介导，尽管大脑的灵敏度的基础 尚不清楚钙调蛋白的循环酶。 只有一张新兴的图片 激素受体 - 腺苷酸环化酶系统的成分数量， 没有清楚地了解其互动机制 调节循环酶活性。 拟议的研究重点是钙调蛋白诱导的机制 腺苷酸环化酶的激活，并将确定它是否：a）涉及 刺激和抑制性鸟嘌呤核苷酸的参与 监管组件； b）涉及其他未知组件，独有的 钙调蛋白敏感酶； c）居住在独特的特性中 钙调蛋白敏感酶的催化单位；或d）涉及一个 这些可能性的结合。 两种主要方法将是 净化Ca2+/钙调蛋白敏感腺苷酸环化酶并识别 带有标记的钙调蛋白和交联剂钙调蛋白结合 与该酶相关的蛋白质。 NS和/或Ni的潜在作用 在钙调蛋白激活时，将通过间接确定 以及直接评估以及通过重组研究。 鉴于 二价阳离子在调节腺苷的调节中起着确定的作用 循环酶活性，其参与钙调蛋白激活的特征 将确定脑酶的。 将准备多克隆和/或单克隆抗体 环化酶催化单元和/或相关的钙调蛋白结合蛋白。 这些将被使用：a）开发免疫力治疗吸附剂；和b）\ to 澄清这种形式的环化酶本地化的问题或 相关的钙调蛋白结合蛋白。 从狭窄的意义上讲，研究将有助于我们理解 单个代理改变腺苷酸环化酶活性，但在更广泛的 Sense将深入了解细胞调节过程的整合 涉及营地和钙。