Molecular signal transduction of cAMP compartments

cAMP 区室的分子信号转导

• 批准号: 8991494
• 负责人: RENNOLDS S OSTROM
• 金额: $ 15.13万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8991494
• 关键词: A kinase anchoring protein; Adenylate Cyclase; Asthma; Biochemical; Biological Assay; Biological Models; Cell Shape; Cell membrane; Cell model; Cell physiology; Cells; Chronic Obstructive Airway Disease; Congestive Heart Failure; Coronary Arteriosclerosis; Coupling; Cyclic AMP; Cyclic AMP Receptors; Data; Disease; Enzymes; Fibroblasts; Fluorescence Resonance Energy Transfer; Future; G-Protein-Coupled Receptors; Goals; Hormone Receptor; Human; Hypertension; Individual; Interleukin-6; Kinetics; Lead; Learning; Life; Location; Membrane Microdomains; Molecular; Monitor; Nature; Neurotransmitter Receptor; Physiological; Positioning Attribute; Production; Protein Isoforms; Proteins; Pulmonary Fibrosis; Receptor Signaling; Role; Second Messenger Systems; Sensitivity and Specificity; Signal Transduction; Smooth Muscle Myocytes; Somatostatin; Source; Stroke; Study models; Therapeutic; Time; base; cell type; design; differential expression; interdisciplinary approach; knock-down; mutant; novel; novel strategies; overexpression; phosphoric diester hydrolase; public health relevance; receptor; receptor coupling; respiratory smooth muscle; response; second messenger; sensor; time use; tool

A large number of G protein coupled receptors (GPCR) utilize cAMP as their second messenger to induce alterations in cell function. In fact, in the same cell several different GPCR can increase cAMP, leading to the question of how the cell interprets the signals from these receptors differently. The concept of cAMP compartmentation, where the second messenger is not generated uniformly throughout the cell, is readily accepted yet poorly understood. We have found that the enzymes that synthesize cAMP, adenylyl cyclases (AC's), are not uniformly distributed through the plasma membrane. Furthermore, GPCR can preferentially couple to certain AC isoforms due to colocalization in lipid rafts. While we have made progress in understanding how specific receptors can couple to certain AC's, little progress has been made in defining the compartments of cAMP inside cells and even less is known about what cellular responses can be modified by different pools of cAMP. One problem has been that common cell models used in the field lack highly compartmentized cAMP pools. We have found that cultured human airway smooth muscle (HASM) cells express identifiable cAMP compartments. Furthermore, we can define these compartments based on the isoforms of AC they express. We have observed that signaling by cAMP generated by AC2, but not AC6 or other AC's, leads to the expression of IL-6 by HASM. Moreover, cAMP generated by AC6, but not by AC2, increases expression of somatostatin and stimulates a cell shape change called arborization. Thus, AC2- and AC6-specific responses can be used to define the cAMP signaling compartments in HASM. The goal of this project is to characterize the other components of these two cAMP compartments by using overexpression and knockdown of specific AKAP's and PDE's. Novel AC mutants will be used to manipulate AC localization and function to determine how these pools are assembled. This project proposes novel, multidisciplinary approaches to define the components responsible for establishing and maintaining cAMP signaling compartments. Results will have broad applicability due to the fundamental nature of cAMP signaling, but because a well-differentiated cell model is used, our findings will also have direct relevance to asthma and COPD therapy. GM107094 Ostrom, Rennolds S 2.

大量G蛋白偶联受体(GPCR)利用cAMP作为其第二信使 引起细胞功能的改变。事实上，在同一细胞中，几种不同的GPCR可以增加cAMP， 这就引出了一个问题，即细胞如何以不同的方式解释来自这些受体的信号。这个 阵营划分的概念，其中第二信使不是统一产生的 在整个细胞中，人们很容易接受，但对此知之甚少。我们已经发现，这些酶 合成cAMP的腺酰环化酶(AC)在血浆中的分布并不均匀 薄膜。此外，由于共定位，gpr可以优先偶联到某些AC异构体 在脂筏上。虽然我们在了解特定受体如何与 某些AC，在确定细胞内cAMP的隔间方面进展甚微 关于不同的cAMP池可以改变什么细胞反应，更是知之甚少。一 问题是，在战场上使用的普通电池模型缺乏高度分隔的营地池。 我们发现，培养的人呼吸道平滑肌(HASM)细胞表达可识别的cAMP 车厢。此外，我们可以根据AC的异构体来定义这些隔室 快递。 我们观察到，由AC2产生的cAMP信号，而不是AC6或其他AC产生的cAMP信号，导致 HASM表达IL-6的实验研究此外，AC6而不是AC2产生的cAMP增加 表达生长抑素，并刺激称为树枝状的细胞形状变化。因此，AC2-和 AC6特异的反应可以用来定义HASM中的cAMP信令间隔。目标是 该项目的目的是通过使用以下方法来表征这两个营区的其他部件 特定AKAP和PDE的过表达和敲除。新型AC突变体将用于 控制交流本地化和功能以确定这些电池组是如何组装的。这个项目 提出新的、多学科的方法来定义负责建立 以及维护军营信号舱。结果将具有广泛的适用性，因为 CAMP信号的基本性质，但因为使用了分化良好的细胞模型，我们的 研究结果还将与哮喘和COPD治疗直接相关。 雷诺兹·S的GM107094奥斯特罗姆 2.