ROLE AND REGULATION OF ENDOGENOUS RGS IN HUMAN CELLS

内源性 RGS 在人体细胞中的作用和调节

• 批准号: 2910040
• 负责人: ANDREJS M KRUMINS
• 金额: $ 3.84万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-05-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2910040
• 关键词: G protein; RNase protection assay; biological signal transduction; cell line; enzyme activity; guanosinetriphosphatase activating protein; guanylate cyclase; protein localization; protein structure function; tissue /cell culture

Understanding mechanisms of regulation of G protein-mediated transmembrane signaling is critical to comprehension and prediction of the effects of hormones, neurotransmitters, and related drugs on cellular function. A large a novel family of negative regulators of G protein function has been discovered recently [1] termed RGS proteins (Regulators of G protein Signaling), these proteins act as powerful GTPase-activating proteins (GAPs) for several heterodimeric G proteins, increasing their rate of GTPase hydrolysis more than 40-fold [2]. RGS proteins act catalytically and are thus likely to play a significant role in attenuation or down regulation of receptor-effector systems where G proteins act as intermediary transducers. I propose to examine the role and regulation of RGS proteins in vivo using a two-tiered approach. The first tier employs RNase protection assays to comprehensively catalog the identity and conditions required for RGS expression. The information obtained from this approach will subsequently be used for the second tier of study; namely, to examine the regulation of RGS protein activity in vivo. RGS proteins identified in the first tier will be tested for expression levels, sub- cellular localization, and post-translational status (e.g. phosphorylation, lipidation and association with regulatory proteins) and correlated with in vitro GAP assay activity. and in vivo effects on G/s, G/i, and G/q coupled receptor systems. RGS expression and activity will be compared in the presence and absence of appropriate hormones and neurotransmitters..

了解G蛋白介导的跨膜调节的机理 信号对理解和预测至关重要 激素，神经递质和相关药物有关细胞功能。一个 G蛋白功能的负调节剂的新型家族已经是 最近发现[1]称为RGS蛋白（G蛋白的调节剂 信号传导），这些蛋白充当强大的GTPase激活蛋白 （差距）几种异二聚体G蛋白，增加了其速率 GTPase水解超过40倍[2]。 RGS蛋白质作用催化作用 因此，很可能在衰减或下降中发挥重要作用 G蛋白充当受体效应系统的调节 中间传感器。我建议检查 RGS蛋白在体内使用两层方法。第一层雇用 RNase保护测定法以全面分类身份和 RGS表达所需的条件。从此获得的信息 随后将使用方法进行第二层研究；即， 检查体内RGS蛋白活性的调节。 RGS蛋白 在第一层中确定的表达水平将测试 细胞定位和翻译后状态（例如 磷酸化，脂化和与调节蛋白的关联）和 与体外间隙测定活性相关。和体内对g/s的影响， G/I和G/Q耦合受体系统。 RGS表达和活动将是 在存在和不存在适当的激素和 神经递质..