Regulation of Adenylyl Cyclase Signaling Pathways

摘要 腺酰环化酶(AC)及其产物环磷酸腺苷(CAMP)调节每种哺乳动物组织的功能结果 和器官系统，控制学习和记忆，运动协调，心脏 收缩、药物依赖和戒断、肾功能、疼痛、应激、免疫反应和焦虑 行为举止，仅举几例。重要的是，G蛋白偶联受体通过刺激传递信号 (GαS)或AC的抑制(GαI)是主要的临床药物靶点(例如治疗心脏病的β-受体阻滞剂，治疗心脏病的阿片类药物 止痛药或治疗哮喘的β-激动剂)。主要由NIGMS资助，在过去的27年里，我的研究生涯 重点阐述了G蛋白等对AC酶催化活性的调节作用 分子。此外，我们感兴趣的是AC/cAMP信号特异性如何发生在细胞内 环境。我们已经鉴定了由A-激酶锚定组装的多个大分子复合体 当AC锚定到系统时，对低水平的局部cAMP产生做出反应的蛋白质，驱动 生理反应，或当改变时，会产生病理生理后果。然而，许多人 关于这些信号体中交流调节的机制，基本的问题仍然存在 通过共价修饰调节ACS，以及从细胞内调节细胞事件的可能性 网站。为了解决这些关键问题，我们执行了近邻的邻近识别 (BioID)在心肌细胞中建立全面的AC相互作用网络并鉴定新的工具 选择性地探测AC9的性质。这些将应用于检查脚手架的广泛项目 最终控制心脏起搏和传导的AC活动。Popdc蛋白作为一种新的 促进复合体内AC活性的AC酶的支架，以驱动cAMP依赖的调节 迷航频道。Popdc调节交流的独特机制和该复合体的定位将是 在两个相关项目中进一步探讨。此外，我们将研究交流调节的机制，通过 新型修饰酶，经BioID鉴定。尽管这些调节cAMP信号的模式将是 在心肌细胞和心脏功能的背景下进行研究，许多机制在 并对许多生物系统产生影响。

Summary Adenylyl cyclase (AC) and its product, cyclic AMP, regulate functional outcomes in every mammalian tissue and organ system, controlling processes such as learning and memory, motor coordination, cardiac contractility, drug dependency and withdrawal, renal function, pain, stress, immune responses, and anxiety behavior, to name just a few. Importantly, G protein coupled receptors that transmit signals via stimulation (Gαs) or inhibition (Gαi) of AC are major clinical drug targets (e.g. beta-blockers for heart disease, opioids for pain, or beta-agonists for asthma). Funded largely by NIGMS, my research career over the last 27 years has focused on how the catalytic activity of AC enzymes are regulated by G proteins and other regulatory molecules. Additionally, we are interested in how AC/cAMP signaling specificity can occur in a cellular environment. We have identified multiple macromolecular complexes assembled by A-kinase anchoring proteins that respond to low levels of local cAMP production upon anchoring of AC to the system, driving physiological responses, or when altered, have pathophysiological consequences. However, many fundamental questions remain about the mechanisms of AC regulation within these signalosomes, the regulation of ACs by covalent modifications, and the potential for regulation of cellular events from intracellular sites. To address these key questions, we have performed proximity dependent identification of near neighbors (BioID) to build a comprehensive AC interaction network in cardiomyocytes and characterized new tools for selectively probing the properties of AC9. These will be applied to broad projects that examine the scaffolding of AC activity that ultimately control cardiac pacemaking and conduction. Popdc proteins act as one novel scaffold for AC enzymes that promote AC activity within the complex to drive cAMP-dependent regulation of TREK channels. The unique mechanism of AC regulation by Popdc and the localization of this complex will be further explored in two related projects. Additionally, we will examine the mechanism of AC regulation by a novel modifying enzyme, identified by BioID. Although these modes of regulating cAMP signaling will be examined in the context of cardiomyocytes and cardiac function, many of the mechanisms are universal in nature and have implications for many biological systems.