Regulation Of Adenylyl Cyclase Signaling Pathways

描述（由申请人提供）：cAMP生产的调控需要一系列复杂的信号分子，这些信号分子目前被用作治疗心脏病、高血压、精神分裂症、帕金森病、哮喘、慢性疼痛等疾病的药物干预靶点。尽管在一系列重要的生理过程和病理生理条件中cAMP是一个基本的“第二信使”，但控制cAMP作用特异性和时间方面的分子机制尚不清楚。被称为akap的一个庞大而复杂的蛋白质家族的发现可能在这种调节中起主要作用。akap最初被认为是将cAMP依赖性蛋白激酶A （PKA）锚定到cAMP/PKA作用的下游靶点。然而，在最近的研究中，我们发现产生cAMP的酶腺苷酸环化酶（AC）的几种异构体也在大脑和心脏中与几种akap复合物中发现，这表明cAMP的产生及其下游靶点是共定位的。此外，我们已经发现AC的上游调节剂异三聚体G蛋白也是AC复合物的一部分，通过它们与AC上先前未被识别的位点结合。本应用旨在了解含有AC的大分子复合物如何引起camp控制的下游事件的动态和特异性调节，例如参与炎症性疼痛的离子通道和调节海马突触活性的谷氨酸受体。三个特定的目的是为了解决包含ACs的信号复合物是camp依赖过程的空间和时间调节所必需的假设。Aim 1将建立AC和异三聚体G蛋白预形成复合物的功能，Aim 2将确定AKAP如何调节AC活性和动力学，Aim 3将确定结合AC对AKAP功能的要求。

DESCRIPTION (provided by applicant): Regulation of cAMP production requires an elaborate series of signaling molecules that are currently used as targets for drug intervention in the treatment of heart disease, hypertension, Schizophrenia, Parkinson's, asthma, chronic pain, and many more. Despite being a fundamental "second messenger" in a huge array of important physiological processes and pathophysiological conditions, the molecular mechanisms that control the specificity and temporal aspects of cAMP actions are not well understood. The discovery of a large and complex family of proteins termed AKAPs likely play a major role in this regulation. AKAPs had originally been thought of as anchoring the cAMP-dependent protein kinase A (PKA), to downstream targets of cAMP/PKA actions. However, in recent studies we have found that several isoforms of the enzyme that produces cAMP, adenylyl cyclase (AC) are also found in complex with several AKAPs in both brain and heart, suggesting that the production of cAMP as well its downstream targets are co-localized. Further, we have found evidence that the upstream regulators of AC, heterotrimeric G proteins, are also a part of an AC complex through their binding to previously unrecognized sites on AC. This application seeks to understand how macromolecular complexes containing AC gives rise to dynamic and specific regulation of cAMP-controlled downstream events, such as ion channels involved in inflammatory pain and glutamate receptors modulating hippocampal synaptic activity. Three specific aims are designed to address the hypothesis that, signaling complexes containing ACs are required for spatial and temporal regulation of cAMP-dependent processes. Aim 1 will establish the function of pre-formed complexes of AC and heterotrimeric G proteins, Aim 2 will determine how AKAPs regulate AC activity and dynamics, and Aim 3 will determine the requirement of bound AC for AKAP function. PUBLIC HEALTH RELEVANCE: Regulation of cAMP production requires an elaborate series of signaling molecules that are currently used as targets for drug intervention in the treatment of heart disease, hypertension, Schizophrenia, Parkinson's, asthma, chronic pain, and many more. Despite being a fundamental signaling molecule in physiological and pathophysiological conditions, the molecular mechanisms that control the specificity and temporal aspects of cAMP actions are not completely understood. This is particularly true for the enzyme that produces cAMP, adenylyl cyclase (AC). We have identified novel multi-protein complexes containing AC that control cAMP signaling in both brain and heart. The existence of these complexes suggests that the production of cAMP as well its downstream targets are co- localized. In addition, the combinations of specific AC isoforms and targets of cAMP actions are far more organized than previously appreciated. This application seeks to understand how macromolecular complexes containing AC gives rise to cAMP regulation and specificity, and to investigate the roles of these complexes in controlling processes involved in inflammatory pain and hippocampal synaptic regulation.