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: 8846612
• 负责人: Alan V. Smrcka
• 金额: $ 30.29万
• 依托单位: UNIVERSITY OF ROCHESTER
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8846612
• 关键词: ADRBK1 gene; Absence of pain sensation; Adenylate Cyclase; Adrenergic Receptor; Affinity; Amino Acids; Animal Model; Binding; Binding Sites; Biological Assay; Biological Models; Cell Line; Cell model; Cells; Complement Receptor; Complex; Coupled; Crystallization; Crystallography; Cyclic AMP; Data; Development; Disease; Family; Funding; G-Protein-Coupled Receptors; G-protein Beta gamma; GTP-Binding Proteins; Heart failure; Hot Spot; Inflammation; Investigation; Laboratories; Lead; Ligands; Link; MAPK3 gene; Measures; Mediating; Modification; Morphine; Mutagenesis; Mutation; NMR Spectroscopy; Pharmacologic Substance; Phosphorylation; Play; Process; Production; Protein Subunits; Proteins; Proto-Oncogene Proteins c-akt; Publishing; Purinoceptor; Receptor Mediated Signal Transduction; Resistance; Role; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Specificity; Structure; Surface; Surface Plasmon Resonance; System; Testing; Therapeutic; Validation; Work; X-Ray Crystallography; base; cofactor; gallein; inhibitor/antagonist; knock-down; macrophage; mutant; novel; novel therapeutic intervention; novel therapeutics; protein protein interaction; receptor; research study; signal processing; small molecule

DESCRIPTION (provided by applicant): G proteins ¿? subunits play a central role in G-protein coupled receptor (GPCR)-mediated signal transduction. They act as cofactors in the receptor-mediated activation process as well as playing direct roles in signal transfer to downstream targets. Considerable data has accumulated in number of systems that excess ¿? signaling has pathological consequences and that manipulation of ¿? subunit signaling could be an effective therapeutic strategy in heart failure as well as other diseases. We developed a novel targeting strategy for selective manipulation of G protein ¿? subunit signaling pathways by selectively blocking ¿? -subunit binding interactions with functional protein partners using small molecules. In the previous funding period we defined the binding modes for several compounds by surface plasmon resonance (SPR) coupled with site directed mutagenesis and solved the co-crystal structure of M201 bound to the hot spot of G¿?. These data confirmed a direct mechanism for binding to G¿? that influences protein-protein interactions and support our overall hypothesis that small molecules selectively modulate downstream effectors signaling by binding to different subsites on the G¿? hotspot. Additionally, we published results demonstrating efficacy and specificity of these compounds in cellular and animal models of heart failure, inflammation and morphine- dependent analgesia. In the experiments proposed in this application we will continue to explore the fundamental mechanisms underlying binding and selectivity of these ¿? binding compounds. Specific aim 1 will focus on mutagenesis and x-ray crystallography to identify multiple binding modes within the G¿? hotspot that contribute to selectivity. Specific aim 2 will explore the mechanism for compound-dependent G¿? subunit activation. Specific aim 3 will explore specificity and mechanism of action in intact cells. Successful completion of the proposed experiments will lead to a thorough understanding of a the mechanism of action of a new family of molecules that target G23 signaling that have potential uses in dissecting the mechanisms of action of GPCR stimulated signaling and providing the basis for novel therapeutic approaches.

描述（由申请人提供）：G 蛋白 ¿亚基在 G 蛋白偶联受体 (GPCR) 介导的信号转导中发挥核心作用。它们在受体介导的激活过程中充当辅助因子，并在信号传递至下游靶点中发挥直接作用。大量系统中已积累了大量数据，超过 ??信号传导具有病理后果，并且操纵??？亚基信号传导可能是心力衰竭和其他疾病的有效治疗策略。我们开发了一种选择性操纵 G 蛋白的新型靶向策略 ??通过选择性阻断 ?? 亚基信号通路-使用小分子与功能性蛋白质伴侣的亚基结合相互作用。在之前的资助期间，我们通过表面等离子共振（SPR）结合定点诱变定义了几种化合物的结合模式，并解决了M201与G??热点结合的共晶结构。这些数据证实了与 G? 结合的直接机制。影响蛋白质-蛋白质相互作用并支持我们的总体假设，即小分子通过结合到 G 上的不同亚位点选择性地调节下游效应器信号传导。热点。此外，我们发表的结果证明了这些化合物在心力衰竭、炎症和吗啡依赖性镇痛的细胞和动物模型中的功效和特异性。在本申请提出的实验中，我们将继续探索这些物质的结合和选择性的基本机制。结合化合物。具体目标 1 将重点关注诱变和 X 射线晶体学，以确定 G¿？有助于选择性的热点。具体目标2将探索化合物依赖性G?的机制。亚基激活。具体目标 3 将探索完整细胞的特异性和作用机制。成功完成所提出的实验将有助于彻底了解以 G23 信号传导为目标的新分子家族的作用机制，这些分子在剖析 GPCR 刺激信号传导的作用机制方面具有潜在用途，并为新的治疗方法提供基础。