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: 8444398
• 负责人: Alan V. Smrcka
• 金额: $ 29.23万
• 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/8444398
• 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.

描述（由应用程序提供）：G蛋白质�？亚基在G蛋白偶联受体（GPCR）介导的信号转移中起着核心作用。它们在受体介导的激活过程中充当辅助因子，并在信号传递到下游目标中扮演直接作用。超出超过�的系统数量积累了大量数据？信号传导具有病理后果和对„的操纵？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。我们制定了一种新颖的靶向策略来选择性操纵G蛋白�？亚基信号通路通过选择性阻止�？ - 使用小分子与功能蛋白伴侣的subumunit结合相互作用。在上一个资金期间，我们通过表面等离子体共振（SPR）与位点的定向诱变定义了几种化合物的结合模式，并求解了M201的共结晶结构，结合到G？的热点。这些数据证实了与G？结合的直接机制？这影响了蛋白质 - 蛋白质相互作用，并支持我们的总体假设，即小分子通过与G g上的不同亚矿石结合选择性调节下游效应信号传导？热点。此外，我们发表了结果，证明了这些化合物在心力衰竭，感染和吗啡依赖性镇痛的细胞和动物模型中的有效性和特异性。在本应用程序中提出的实验中，我们将继续探索这些应用程序的基本机制和选择性的基本机制？结合化合物。特定的目标1将重点放在诱变和X射线晶体学上，以识别g g。内的多种结合模式？有助于选择性的热点。具体目标2将探索复合依赖性g¿的机制？亚基激活。特定的目标3将探索完整细胞中的特异性和作用机理。成功完成所提出的实验将导致对靶向G23信号的新分子家族的作用机理，该作用机理在解剖GPCR刺激信号传导的作用机理方面具有潜在用途，并为新型治疗方法提供了基础。