GABA TRANSPORTER INTERACTING DOMAINS

GABA 转运蛋白相互作用域

• 批准号: 6660680
• 负责人: MICHAEL W. QUICK
• 金额: $ 24.38万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-05-10 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6660680
• 关键词: animal tissue; biological signal transduction; chemical association; chloride channels; density gradient ultracentrifugation; gamma aminobutyrate; gene deletion mutation; gene expression; glia; immunoprecipitation; intermolecular interaction; membrane transport proteins; mutant; neurons; neuroregulation; neurotransmitter transport; protein binding; protein folding; protein protein interaction; protein structure function; syntaxin; tissue /cell culture

DESCRIPTION: (Applicant's Abstract) Neurotransmitter levels in brain are critical for normal brain function. Abnormal levels of neurotransmitter, resulting in inappropriate neural signaling, underlie a diverse list of brain disorders. For example, epilepsy, excitotoxic cell death, depression, and a number of conditions related to drug abuse are all related to abnormal transmitter levels in brain. Neurotransmitter transporters are proteins, located on neurons and glia, that function in part to transport transmitter from the extracellular milieu into cells. As such, they play a central role in regulating synaptic signaling. While much is being learned about the permeation properties of various neurotransmitter transporters (e.g., transport rates, substrate affinities, conducting states), little is known about how particular structural domains of the transporter participate in the permeation process. Furthermore, while it is known that transporter function can be regulated, the extent to which the regulation occurs through manipulation of the domains involved in permeation is not known. The major goal of this application is to define the role of intra-molecular and inter-molecular interactions in regulation of the GABA transporter GAT1. Specific Aim 1 is to test the hypothesis that the amino terminal cytoplasmic tail of the GABA transporter GAT1 positively regulates GABA transport through interactions with internal cytoplasmic loops of the transporter. Specific Aim 2 is to test the hypothesis that proteins that physically interact with the amino terminal tail of GAT1 negatively regulate GAT1 function by preventing the N-terminal tail from interacting with the internal cytoplasmic loops of the transporter. Specific Aim 3 is to test the hypothesis that intermolecular interactions that negatively regulate transporter function can be modulated by physiologically relevant factors. These studies are important because they will (i) define a new regulatory role in permeation for intracellular transporter domains; (ii) elucidate a novel mechanism for the cellular regulation of transporter function through protein-protein interactions; and (iii) provide data that could be useful in strategies aimed at regulating transporter function in the treatment of disorders related to abnormal transmitter levels.

描述：（申请人摘要） 大脑中的神经递质水平对于正常的大脑功能至关重要。 神经递质水平异常，导致不适当的神经 信号传导是多种大脑疾病的基础。例如，癫痫症、 兴奋性毒性细胞死亡、抑郁症以及许多与药物相关的病症 滥用都与大脑中异常的递质水平有关。神经递质 转运蛋白是位于神经元和神经胶质细胞上的蛋白质，其部分功能 将递质从细胞外环境转运到细胞内。像这样， 它们在调节突触信号传导中发挥核心作用。虽然很多事情正在发生 了解各种神经递质的渗透特性 转运蛋白（例如转运速率、底物亲和力、导电状态）， 关于转运蛋白的特定结构域如何 参与渗透过程。此外，虽然已知 转运体功能可以被调节，调节的程度 通过操纵涉及渗透的域而发生的情况尚不清楚。 该应用的主要目标是定义分子内和 GABA 转运蛋白 GAT1 调节中的分子间相互作用。 具体目标 1 是检验氨基末端细胞质的假设 GABA 转运蛋白 GAT1 的尾部通过以下方式正向调节 GABA 转运： 与转运蛋白内部细胞质环的相互作用。具体目标2 是为了检验以下假设：蛋白质与氨基发生物理相互作用 GAT1 的末端尾部通过阻止 N 末端尾巴与内部细胞质环相互作用 运输者。具体目标 3 是检验分子间相互作用的假设 负调节转运蛋白功能的相互作用可以通过以下方式调节 生理相关因素。 这些研究很重要，因为它们将 (i) 定义新的监管角色 细胞内转运蛋白结构域的渗透； (ii) 阐明一部小说 转运蛋白功能的细胞调节机制 蛋白质-蛋白质相互作用； (iii) 提供可能有用的数据 旨在调节转运蛋白功能的治疗策略 与异常递质水平相关的疾病。