Selective Targeting of G Protein beta gamma Subunits with Small Molecules

小分子选择性靶向 G 蛋白 β γ 亚基

• 批准号: 9321302
• 负责人: Alan V. Smrcka
• 金额: $ 30.4万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9321302
• 关键词: Active Sites; Adhesions; Binding; Biochemical; Biological Assay; Biology; Cell Adhesion; Cell Polarity; Cell membrane; Cell physiology; Cells; Cellular biology; Chemotactic Factors; Co-Immunoprecipitations; Complement; Coupling; Cyclic AMP; Development; Disease; Drug Targeting; Funding; G-Protein-Coupled Receptors; G-protein Beta gamma; G-substrate; GTP-Binding Protein alpha Subunits; GTP-Binding Proteins; HL60; Heterotrimeric G Protein Subunit; Heterotrimeric GTP-Binding Proteins; Hormones; Immobilization; Immune; In Vitro; Individual; Integrins; Investigation; Laboratories; Lead; Ligands; Light; Lighting; Mammalian Cell; Measures; Mediating; Modification; Molecular; Molecular Target; Neurotransmitters; Normal Cell; Optics; Pathway interactions; Pharmacologic Substance; Protein Analysis; Protein Subunits; Receptor Signaling; Recruitment Activity; Regulation; Role; Second Messenger Systems; Series; Shapes; Signal Pathway; Signal Transduction; Site; Specificity; Talin; Therapeutic; cell behavior; cell motility; chemokine; chemokine receptor; directional cell; experimental study; fMet-Leu-Phe receptor; fibrosarcoma; gallein; inhibitor/antagonist; migration; neutrophil; novel; novel therapeutics; overexpression; polarized cell; protein activation; receptor; receptor coupling; response; small molecule; small molecule inhibitor; spatiotemporal; targeted treatment; therapeutic target; tool

Abstract G protein-coupled receptors (GPCRs) mediate the actions of a wide variety of hormones and neurotransmitters to control functions in all mammalian cells. As such, GPCRs are major targets of therapeutics. Individual GPCRs directly couple to distinct complements of heterotrimeric G protein  and  subunits that drive downstream signaling pathways to shape the cellular responses that determine GPCR efficacy. Both G and G subunits interact directly with effectors to produce cellular responses. Our laboratory has focused largely on G subunit signaling and has identified a series of small molecules that bind to G to “bias” signaling pathways downstream of GPCRs. These inhibitors have been applied extensively to dissect G functions in biology, and to validate G as a viable therapeutic target. Most recently we have identified a small molecule that activates G subunit signaling without activating G protein  subunits. We have applied this tool to study signaling in immune cells to reveal exciting and previously unappreciated roles for Gi in chemokine- chemoattractant receptor signaling. In this proposal we will examine in detail how G activating molecules function at a molecular level, and explore the mechanistic role for Gi in chemoattractant receptor signaling in the following specific aims. Aim 1. Determination of the mechanisms of action of 12155 and M119/gallein on G protein subunit interactions and GPCR coupling. Aim 2. Determining the mechanism(s) for GiGTP- dependent regulation of cell migration and adhesion. Aim 3. Analysis of spatiotemporal regulation of Gi activation in chemoattractant-dependent directional cell migration. We believe that these studies will reveal a new molecular target and cellular function for Gi in regulation of integrin function that will have wide implications for chemoattractant/chemokine receptor and integrin regulation.

抽象的 G 蛋白偶联受体 (GPCR) 介导多种激素和神经递质的作用 控制所有哺乳动物细胞的功能因此，GPCR 是个体治疗的主要目标。 GPCR 直接与异源三聚体 G 蛋白 α 和 β 亚基的不同互补体偶联，驱动 下游信号通路塑造决定 GPCR 功效的细胞反应。 G 亚基直接与效应器相互作用以产生细胞反应。 研究人员对 G 亚基信号传导进行了研究，并鉴定了一系列与 G 结合以产生“偏向”信号传导的小分子 这些抑制剂已广泛应用于剖析 G 的功能。 生物学，并验证 G 作为可行的治疗靶点。最近我们发现了一种小分子。 激活 G 亚基信号传导而不激活 G 蛋白  亚基我们已应用该工具进行研究。 免疫细胞中的信号传导揭示了 Gαi 在趋化因子中令人兴奋且以前未被认识到的作用 在本提案中，我们将详细研究 G 激活分子的机制。 在分子水平上发挥作用，并探讨 Gαi 在趋化受体信号传导中的机制作用 目标 1. 确定 12155 和 M119/gallein 对 G 的作用机制。 蛋白质亚基相互作用和 GPCR 偶联 目标 2. 确定 GiGTP- 的机制。 细胞迁移和粘附的依赖性调节 目标 3. Gi 的时空调节分析。 我们相信这些研究将揭示趋化剂依赖性定向细胞迁移的激活。 Gi 调节整合素功能的新分子靶点和细胞功能将具有广泛的应用前景 对趋化剂/趋化因子受体和整合素调节的影响。