MOLECULAR BASIS OF SIGNAL TRANSDUCTION

信号转导的分子基础

• 批准号: 2177345
• 负责人: MELVIN SIMON
• 金额: $ 30.28万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2177345
• 关键词: G protein; cell differentiation; embryonic stem cell; gene expression; genetically modified animals; laboratory mouse; molecular cloning; nucleic acid sequence; phospholipase C; protein isoforms; protein structure; receptor; receptor coupling; tissue /cell culture; transfection

Heterotrimeric G-proteins mediate the activity of hundreds of different serpentine receptors (seven pass membrane receptors). They couple the ligand binding event at the extracellular surface to the activity of an intracellular or membrane bound effector. Thus, they play a critical role in an enormous number of physiological processes including sensory perception, hormone action, neurotransmission, the inflammatory response, and perhaps in growth control and morphogenesis. A clear understanding of how they function could result in the development of drugs that block or enhance these processes. Our goals are to analyze the diversity in the heterotrimeric G-protein family and to determine how G proteins function to generate the appropriate connections between receptors and effectors with the correct kinetics to regulate function. The heterotrimeric G proteins are composed of a large family of genes that encode multiple alpha, beta, and gamma subunits. Our hypothesis is that the combinatorial association of these components leads to the formation of a large variety of heterotrimers embodying specificity for different receptors and effectors. In this renewal we propose to: 1. Further explore the diversity of the Galpha, beta, and gamma subunit genes. 2. Study the basis for isoform specific interaction between members of the Gq family and specific phospholipase C isozymes using transient transfection, mutagenesis, and in vitro reconstitution assays. 3. Search for the receptors and effectors that couple to the Galpha12 family of G proteins. 4. Explore the basis of the cell type specificity of expression of certain G proteins. 5. Target the genes encoding the Gq and G(12) family of alpha subunits using homologous recombination in embryonic stem cells (ES cells) and eliminate these gene functions. We will generate mice that lack polypeptides corresponding to specific Galpha and Ggamma subunits. These animals and the ES cell lines will be assayed to determine the effects of the absence of a specific G-protein on physiological and cellular function.

异三聚体G蛋白介导数百种不同的活性 蛇形受体（七个通过膜受体）。 他们夫妇 在细胞外表面的配体结合事件与 细胞内或膜结合效应子。 因此，他们发挥着批判性 在包括感觉的大量生理过程中的作用 感知，激素作用，神经传递，炎症反应， 也许是在生长控制和形态发生中。 清晰的理解 它们的功能如何导致阻塞药物的发展 或增强这些过程。 我们的目标是分析异三聚体G蛋白的多样性 家族并确定G蛋白如何产生生成 受体和效应器之间的适当连接，正确 动力学调节功能。 异三聚体G蛋白由大型基因家族组成 该编码多个alpha，beta和伽马亚基。 我们的假设是 这些组件的组合关联导致 形成了各种各样的异三聚体，体现了特异性的特异性 不同的受体和效应子。 在此续约中，我们建议： 1。进一步探索Galpha，beta和Gamma亚基的多样性 基因。 2。研究同工型特定相互作用的基础 使用瞬态的GQ家族和特定的磷脂酶C同工酶 转染，诱变和体外重构测定法。 3。搜索夫妇到Galpha12的受体和效应子 G蛋白家族。 4。探讨细胞类型表达的基础 某些G蛋白。 5。靶向编码GQ和G（12）α亚基家族的基因 在胚胎干细胞（ES细胞）中使用同源重组和 消除这些基因功能。 我们将产生缺乏的老鼠 对应于特定Galpha和Ggamma亚基的多肽。 这些 动物和ES细胞系将分析以确定影响 没有特定的G蛋白在生理和细胞上 功能。