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: 8234909
• 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 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8234909
• 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; computerized data processing; gallein; inhibitor/antagonist; knock-down; macrophage; mutant; novel; novel therapeutic intervention; novel therapeutics; protein protein interaction; receptor; research study; 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. PUBLIC HEALTH RELEVANCE: G protein coupled receptors (GPCRs) are a major class of transmembrane receptors responsible for recognition of a large class of diverse ligands. Here we propose investigation of selective small molecule inhibitors of G protein ¿? subunits identified in our laboratory which could be used to inhibit multiple GPCRs and modify actions of existing GPCR directed pharmaceuticals. Results of these experiments will help to validate this alternate approach to modification of signaling pathways downstream of GPCRs that could ultimately lead to development of novel therapeutics.

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着核心作用，它们在受体介导的激活过程中发挥着辅助因子的作用，并在向下游靶标的信号传递中发挥直接作用。超过 ¿ 的系统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 ¿ -使用小分子与功能蛋白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，并解决了与热结合的M201的共晶结构。 G 点?.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，我们将继续探索结合和选择性的基本机制。其中 ¿ ? 结合化合物。具体目标 1 将重点关注诱变和 X 射线晶体学，以识别 G¿ 内的多种结合模式。 ? 有助于选择性的热点 2 将探索化合物依赖性 G¿特定目标 3 将探索完整细胞中的特异性和作用机制，成功完成拟议的实验将导致对具有潜在用途的靶向 G23 信号传导的新分子家族的作用机制的全面了解。剖析 GPCR 刺激信号传导的作用机制，并为新的治疗方法提供基础。 公共健康相关性：G 蛋白偶联受体 (GPCR) 是一类主要的跨膜受体，负责识别一大类不同的配体。在此，我们建议研究 G 蛋白的选择性小分子抑制剂。我们实验室鉴定出的亚基可用于抑制多种 GPCR 并改变现有 GPCR 导向药物的作用，这些实验结果将有助于验证这种修改 GPCR 下游信号通路的替代方法，最终可能导致新型药物的开发。疗法。