Regulation Of Adenylyl Cyclase Signaling Pathways

描述(申请人提供)：调节cAMP的产生需要一系列精心设计的信号分子，目前这些分子被用作治疗心脏病、高血压、精神分裂症、帕金森病、哮喘、慢性疼痛等药物干预的靶点。尽管在一系列重要的生理过程和病理生理条件下，cAMP是一个基本的“第二信使”，但控制cAMP作用的特异性和时间性的分子机制还不是很清楚。被称为AKAPs的一个庞大而复杂的蛋白质家族的发现可能在这一调控中发挥了重要作用。AKAP最初被认为是将cAMP依赖的蛋白激酶A(PKA)锚定在cAMP/PKA作用的下游靶标上。然而，在最近的研究中，我们发现在大脑和心脏中也发现了产生cAMP的几种同工酶-腺酰环化酶(AC)与几个AKAP的复合体，这表明cAMP的产生及其下游靶点是共定位的。此外，我们还发现证据表明，AC的上游调节因子，异三聚体G蛋白，也是AC复合体的一部分，通过它们与AC上以前未被识别的位点结合。这项应用旨在了解含有AC的大分子复合体如何对cAMP控制的下游事件产生动态和特异性的调节，如参与炎性疼痛的离子通道和调节海马突触活动的谷氨酸受体。设计了三个特定的目标来解决这样的假设，即含有ACS的信号复合体对于cAMP依赖的过程的空间和时间调节是必需的。目标1将确定AC和异源三聚体G蛋白的预先形成的复合体的功能，目标2将确定AKAP如何调节AC的活性和动力学，目标3将确定结合的AC对AKAP功能的需求。 与公共健康相关：调控cAMP的产生需要一系列精心设计的信号分子，目前这些分子被用作治疗心脏病、高血压、精神分裂症、帕金森病、哮喘、慢性疼痛等药物干预的靶点。尽管在生理和病理生理条件下，cAMP是一种基本的信号分子，但控制cAMP作用的特异性和时间方面的分子机制尚不完全清楚。尤其是产生cAMP的酶，腺苷环化酶(AC)。我们已经确定了含有AC的新型多蛋白复合体，它控制着大脑和心脏中的cAMP信号。这些复合体的存在表明cAMP的产生及其下游靶标是共同定位的。此外，特定AC异构体和cAMP作用靶点的组合比以前所认识的要更有组织。本应用旨在了解含有AC的大分子复合体如何引起cAMP调节和特异性，并研究这些复合体在控制炎性疼痛和海马区突触调节过程中的作用。

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.