Regulation of Adenylyl Cyclase Signaling Pathways

总结 腺苷酸环化酶（AC）及其产物环腺苷酸（cAMP）在哺乳动物的每个组织中调节功能结果 和器官系统，控制学习和记忆，运动协调，心脏 收缩性、药物依赖和戒断、肾功能、疼痛、压力、免疫反应和焦虑 行为，仅举几例。重要的是，通过刺激传递信号的G蛋白偶联受体 AC的Gαs或抑制（Gαi）是主要的临床药物靶点（例如，用于心脏病的β受体阻滞剂， 疼痛或哮喘的β-激动剂）。在NIGMS的资助下，我过去27年的研究生涯 重点是AC酶的催化活性如何受到G蛋白和其他调节因子的调节。 分子。此外，我们感兴趣的是AC/cAMP信号特异性如何在细胞内发生， 环境我们已经确定了多个大分子复合物组装的A-激酶锚定 在AC锚定到系统后，响应于低水平的局部cAMP产生的蛋白质， 生理反应，或当改变时，具有病理生理后果。但不少 关于这些信号体中AC调节机制的基本问题仍然存在， 通过共价修饰调节AC，以及从细胞内调节细胞事件的潜力 网站.为了解决这些关键问题，我们已经执行了近邻的邻近依赖识别 （BioID）在心肌细胞中构建全面的AC相互作用网络，并表征了用于 选择性地探测AC 9的性质。这些将被应用到广泛的项目，检查脚手架 最终控制心脏起搏和传导的AC活动。Popdc蛋白作为一种新的 AC酶的支架，其促进复合物内的AC活性以驱动cAMP依赖性调节， TREK频道Popdc的AC调节的独特机制和该复合物的定位将是 在两个相关项目中进一步探索。此外，我们将通过一个 新修饰酶，经BioID鉴定。尽管这些调节cAMP信号传导的模式将被 在心肌细胞和心脏功能的背景下进行检查，许多机制是普遍的， 自然界，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.