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Novel Mechanisms Regulating the Heterotrimeric G Protein Complex

调节异源三聚体 G 蛋白复合物的新机制

基本信息

批准号：
8017477
负责人：
ALAN M. JONES
金额：
$ 33.56万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-01 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8017477
关键词：
Abate Adaptor Signaling Protein Affect Animal Model Apical Arabidopsis Binding Biochemical Biological Assay Biological Models Boxing Caenorhabditis elegans Cell Proliferation Cell Surface Receptors Cell surface Cells Chronic Collaborations Collection Complex Coupled Coupling Data Disease Dissociation Drosophila melanogaster Drug Delivery Systems Elements Engineering Equilibrium Eukaryota Figs - dietary G Protein-Coupled Receptor Genes GTP Binding GTP-Binding Protein Regulators GTP-Binding Proteins Genes Genetic Genetic Screening Germination Glucose Goals Growth Guanine Nucleotide Exchange Factors Guanosine Triphosphate Guanosine Triphosphate Phosphohydrolases Health Heterotrimeric GTP-Binding Proteins Homeostasis Hormones Human Human Engineering Hydrolysis In Vitro Indium Integral Membrane Protein Kinetics Length Ligand Binding Ligands Luciferases Measures Mediating Meristem Methods Modeling Molecular Molecular Genetics Mutation Neurotransmitters Nucleotides Oncogenes Organism Orthologous Gene Pathway interactions Pharmacology Pharmacotherapy Photoaffinity Labels Physiological Physiological Processes Plant Roots Plants Property Proteins Publishing RGS Proteins RGS1 gene Recombinants Regulation Regulatory Element Reporter Genes Research Role Saccharomyces Seeds Signal Pathway Signal Transduction Stem cells Stimulation of Cell Proliferation Structure Testing Transmembrane Domain Uncertainty Work Yeasts base blood glucose regulation cell behavior cell growth comparative dimer driving force drug discovery gene discovery genetic regulatory protein gluconate glucose receptor histogenesis human disease in vivo innovation insight interest meetings mutant novel nucleotide receptor physical property protein complex protein structure prototype receptor research study small molecule sugar tool

项目摘要

DESCRIPTION (provided by applicant): Hormones and neurotransmitters modulate a variety of physiological processes in cell growth and behavior. Their cognate cell surface receptors, which have seven transmembrane (7TM) domains, act by coupling to G proteins, promoting the dissociation of GDP and the subsequent loading of GTP. Signaling abates when GTP is hydrolyzed and GTPase activity is accelerated by Regulators of G Signaling (RGS) proteins having GTPase accelerating protein (GAP) activity. Recently, we discovered a naturally-occurring 7TM-RGS protein in Arabidopsis (AtRGS1) that we hypothesize to be a D-glucose receptor that has a D-glucose-dependent GAP activity. It is the first example of a receptor-GAP and is the prototype for a new class of D-glucose receptors. We also showed that the Arabidopsis G1 subunit has rapid nucleotide exchange making nucleotide hydrolysis the rate limiting step. This property is in marked contrast to the slow nucleotide exchange property of all tested G1 subunits where GDP release is the rate limiting step of the G protein cycle. Thus, we hypothesize that regulation of the G protein cycle is at the GTP hydrolysis step and is mediated by AtRGS1. Finally, we showed that a D-glucose metabolite dramatically increases the nucleotide hydrolysis rate of the G1 subunit. Clearly, the Arabidopsis G protein cycle contains several interesting properties, namely activation of a G1 subunit that does not require a GEF, regulation of the cycle at the GTP hydrolysis step, and a 7TM protein that may be the ligand-regulated GAP controlling the cycling rate. Because the Arabidopsis G1 has the basic core structure and function of human G1, an understanding of how the Arabidopsis G1 activation is regulated will provide insight into novel mechanisms to control human G1 activation. The goal here is to understand how the G1 protein is activated in the context of sugar signaling. Both hypothesis- and discovery-driven approaches will be taken to determine precisely what structure imparts regulatory control. Our initial study of the Arabidopsis G protein cycle illustrated how the G-protein cycle can be regulated by mechanisms apart by the classical GEF. Consequently, a greater degree of plasticity of the cycle is now appreciated and new entry points for regulation are revealed. Understanding the structure underlying these new mechanisms will provide a new means to regulate other G protein cycles. In humans, 7TM receptors and RGS proteins interact either directly or indirectly via adaptor proteins; these are two of several possible mechanisms providing selectivity between receptors and RGS proteins. AtRGS1 is the most extreme example of a mechanism providing receptor-RGS protein selectivity in that both GEF and GAP are two domains on one molecule. Understanding how AtRGS1 regulates the G protein cycle in a ligand dependent manner opens up new possibilities to regulate G protein cycles through drug therapies. Finally, use of Arabidopsis as a model will enable us to solve how cells respond to D-glucose within the context of a multicellular organism. PUBLIC HEALTH RELEVANCE The rationale for the proposed work is that an understanding of molecular mechanisms used in divergent signaling pathways will yield new drug targets, new ideas for manipulating human signaling pathways, and new tools to engineer human pathways.

描述(申请人提供)：荷尔蒙和神经递质调节细胞生长和行为的各种生理过程。它们的同源细胞表面受体有7个跨膜(7TM)结构域，通过与G蛋白偶联而发挥作用，促进GDP的解离和随后的GTP负载。当GTP被水解时，信号转导减弱，GTP酶活性被具有GTP酶加速蛋白(GAP)活性的G信号(RGS)蛋白的调节所加速。最近，我们在拟南芥中发现了一个自然存在的7TM-RGS蛋白(AtRGS1)，我们推测它是一个D-葡萄糖受体，具有D-葡萄糖依赖的GAP活性。这是第一个受体间隙的例子，也是一类新的D-葡萄糖受体的原型。我们还发现，拟南芥G1亚基具有快速的核苷酸交换，使得核苷酸的降解成为速率限制步骤。这一特性与所有测试的G1亚基的缓慢核苷酸交换特性形成鲜明对比，在G1亚基中，GDP的释放是G蛋白周期的速率限制步骤。因此，我们假设G蛋白周期的调节是在GTP水解步骤，并由AtRGS1介导。最后，我们发现D-葡萄糖代谢物显著提高了G1亚基的核苷酸水解率。显然，拟南芥G蛋白周期包含几个有趣的特性，即激活不需要全环基金的G1亚基，在GTP水解步骤调节周期，以及可能是配体调节的间隙控制周期速率的7TM蛋白。由于拟南芥G1具有人类G1的基本核心结构和功能，了解拟南芥G1激活是如何调控的将有助于深入了解控制人类G1激活的新机制。这里的目标是了解在糖信号背景下G1蛋白是如何被激活的。将采取假设驱动和发现驱动的方法来准确确定赋予监管控制的结构是什么。我们对拟南芥G蛋白周期的初步研究表明，G蛋白周期如何通过经典的全球环境基金以外的机制来调节。因此，现在人们认识到了周期的更大程度的可塑性，并揭示了监管的新切入点。了解这些新机制背后的结构将为调控其他G蛋白周期提供新的手段。在人类中，7TM受体和RGS蛋白通过接头蛋白直接或间接相互作用；这是在受体和RGS蛋白之间提供选择性的几种可能机制中的两种。AtRGS1是提供受体-RGS蛋白选择性机制的最极端的例子，因为全球环境基金和GAP都是一个分子上的两个结构域。了解AtRGS1如何以配体依赖的方式调节G蛋白周期，为通过药物治疗调节G蛋白周期开辟了新的可能性。最后，使用拟南芥作为模型将使我们能够在多细胞生物体的背景下解决细胞对D-葡萄糖的反应。公共卫生相关性这项拟议工作的基本原理是，对不同信号通路中使用的分子机制的理解将产生新的药物靶点，操纵人类信号通路的新想法，以及设计人类通路的新工具。