MOLECULAR AND CELLULAR ANALYSIS OF BRAIN ENRICHED PTPS

描述(摘自申请人的摘要)： R01的应用重点是蛋白酪氨酸磷酸酶的作用 (PTPs)在基底节的信号转导通路中的表达。令人信服的证据 提示基底节参与了许多神经精神疾病 精神障碍，包括精神分裂症、抽动症和强迫症 强迫症。在亨廷顿和帕金森氏症中， 基底节，或在它们的传出和传入通路中已被发现。 这种原子核的集合也是许多不想要的东西的活动场所 抗精神病药物的副作用。神经信号转导中断 基底节内的机制是所有这些机制的共同组成部分 紊乱，需要对正常的信号通路有进一步的了解 发生在中南半球的这一区域。 我这项研究的长期目标是确定有助于 神经精神障碍。在以前的筹资机制下(即K20，然后 NIMH一等奖)，我的实验室分离出了一个新的家族或蛋白质 酪氨酸磷酸酶(PTPs)仅存在于大鼠脑内的多巴胺能神经元内 基底节及相关结构。因为蛋白质高度浓缩 在大鼠纹状体中，它被称为STEP。 本申请是我的第一个奖项的自然延续，我们在其中 分离出STEP家族的四种亚型。第一个具体目标是提出 继续隔离这个庞大而重要的群体中具有特征的成员 PTP的数量。该家族由胞质和膜相关的异构体组成，如 以及缺乏活性催化活性的截断形式。其中最 显著的特征是一些步骤包含跨膜结构域，PEST 序列和富含多聚脯氨酸的序列。我们假设这些提供了一种 针对STEP成员的亚细胞靶向机制，用于调节 它们的酶活性，以及它们的底物特异性。此外，我们 认为STEP的磷酸化和蛋白分解发生在多巴胺D1之后 发信号。我们将确定STEP的生理底物，其中之一 被认为是NMDA受体。我们将使用这些假设 生化、分子和电生理技术。

DESCRIPTION (from applicant's abstract): This R01 application focuses on the role of protein tyrosine phosphatases (PTPs) in signal transduction pathways in the basal ganglia. Compeling evidence suggests that the basal ganglia are involved in a number of neuropsychiatric disorders, including schizophrenia, Tourette's syndrome and obsessive compulsive disorder. In Huntington and Parkinson's disorders, abnormalities of the basal ganglia, or in their efferent and afferent pathways have been found. This collection of nuclei are also the site of action for many of the unwanted side effects of neuroleptic drugs. Disruption of the neural signaling mechanisms within the basal ganglia is a common component of all of these disorders, and further knowledge is needed of the normal signaling pathways that occur in this region of the CNS. The long-term goal of my research are to characterize genes that contribute to neuropsychiatric disorders. Under previous funding mechanisams (a K20 and then a FIRST award from NIMH), my laboratory has isolated a novel family or protein tyrosine phosphatases (PTPs) uniquely present within dopaminoceptive neurons of the basal ganglia and related structures. As the protein is highly enriched within rat striatum, it is termed STEP for Striatal Enriched Phosphatase. The present application is a natural continuation of my FIRST Award in which we isolated four isoforms of the STEP family. The first specific aim proposes to continue to isolate the characterize members of this large and important group of PTPs. The family consists of cytosolic and membrane-associated isoforms, as well as truncated forms that lack on active catalytic siste. One of the most striking features is that some STEPs contain transmembrane domains, PEST sequences, and polyproline-rich sequences. We hypothesize that these provide a mechanism for the subcellular targeting of STEP members, for the regulation of their enzymatic activity, and for their substrate specificity. In addition, we propose that phosphorylation asnd proteolysis of STEP occurs after dopamine D1 signaling. We will identify the physiological substrates of STEP, one of which is proposed to be the NMDA receptor. We will these these hypotheses using biochemical, molecular, and electrophysiological techniques.