GABA TRANSPORTER INTERACTING DOMAINS

描述：（申请人摘要） 大脑中的神经递质水平对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，是各种大脑疾病的基础。例如，癫痫， 兴奋性毒性细胞死亡，抑郁症，以及一些与药物相关的疾病 滥用药物都与大脑中不正常的递质水平有关。神经递质 转运蛋白是位于神经元和神经胶质上的蛋白质， 将细胞外环境中的递质转运到细胞内。因此，在本发明中， 它们在调节突触信号传导中起中心作用。虽然很多人 了解了各种神经递质的渗透特性 转运体（例如，传输速率、衬底亲和力、导电状态）， 关于转运蛋白的特定结构域 参与渗透过程。此外，虽然已知 转运蛋白功能可以被调节，调节的程度 通过操纵渗透中涉及的区域而发生的渗透是未知的。 本申请的主要目标是定义分子内和 GABA转运蛋白GAT 1调节中的分子间相互作用。 具体目的1是检验氨基末端细胞质 GABA转运蛋白GAT 1的尾部通过以下途径积极调节GABA转运： 与转运蛋白的内部细胞质环的相互作用。具体目标2 是检验与氨基发生物理相互作用的蛋白质的假设 GAT 1的末端尾部通过阻止GAT 1的表达来负调节GAT 1的功能。 N-末端尾与细胞质内环相互作用， 传送器具体目标3是检验分子间 负调节转运蛋白功能的相互作用可以通过 生理相关因素。 这些研究很重要，因为它们将（i）确定一个新的监管角色 细胞内转运蛋白结构域的渗透;（ii）阐明一种新的 转运蛋白功能的细胞调节机制， 蛋白质-蛋白质相互作用;和（iii）提供可能有用的数据， 治疗中调节转运蛋白功能的策略 与异常的递质水平有关的疾病。

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.