Specifically and regulation TCR pathway protein tyrosine kinases and phosphatases

特异调节TCR通路蛋白酪氨酸激酶和磷酸酶

基本信息

批准号：
8378238
负责人：
ARTHUR WEISS
金额：
$ 89.17万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8378238
关键词：
Affect Behavior Binding Biochemical Biological Biological Models C-terminal Cell membrane Cell model Cells Complex Computer Simulation Discrimination Disease Enzymes Equilibrium Event Feedback Gene Expression Genetic Goals ITAM Kinetics LCP2 gene Lead Lipid Bilayers Lymphocyte-Specific p56LCK Tyrosine Protein Kinase Mediating Membrane Modeling Molecular Mus Noise PTPRC gene Pathway interactions Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Play Property Protein Tyrosine Kinase Protein Tyrosine Phosphatase Proteins Receptor Signaling Recombinant Proteins Recruitment Activity Regulation Regulatory Pathway Role Signal Transduction Site Solutions Specificity Stimulus Study models Surface System T Cell Receptor Signaling Pathway T cell response T cell therapy T-Cell Receptor T-Lymphocyte Tyrosine Tyrosine Phosphorylation Work ZAP-70 Gene analog base expectation inhibitor/antagonist kinase inhibitor membrane model mutant novel pathogen receptor research study response segregation src-Family Kinases two-dimensional

项目摘要

Although most of the molecules involved in TCR signaling have likely been identified, our understanding of the basal signaling versus the induced signaling states are rather rudimentary. Tyrosine kinases and phosphatases maintain the dynamic equilibrium that controls and maintains the both the basal state and changes in their functional activities or spatial localization are important for Induced tyrosine phosphorylation. This project focuses on understanding the specificity and complex regulation of these tyrosine kinases and phosphatases on at the plasma membrane. Understanding the regulation of phosphorylation of the TCR cytoplasmic Z-chain as well as of LAT proteins will be the endpoints we will focus on. We will use: purified recombinant proteins; novel genetically controlled kinase inhibitors; a model two dimensional lipid bilayer system containing defined quantities of recombinant proteins; and, computational modeling to describe and compare the simple bilayer system to existing well-studied intact cekks or membranes of model cellular systems. We will compare the biochemical behavior of such two dimensional models to cellular systems in an iterative way, increasing component complexity and by applying computational modeling. We expect such analyses to reveal expected as well as unanticipated behaviors. We also expect the comparison of the simple system to more complex systems to reveal the presence of regulatory pathways that may not have previously been appreciated, such as positive and negative feedback circuitry. We clearly recognize that we cannot model all of the complexity of TCR signaling but we aim to understand the detailed mechanisms controlling tyrosine phosphorylation of the TCR Z-chain and how these events lead to LAT phosphorylation. The overall goal of project #1 is to understand the specificity and regulation of protein tyrosine kinases and phosphatases involved In TCR signaling that lead to ITAM and LAT phosphorylation at the membrane surface. We will: 1) define the molecular basis for specificity for Lck and ZAP-70 for ITAMs and LAT, respectively; 2) develop an analog sensitive inhibitor system for Lck; 3) define the mechanisms that control ITAM phosphorylation by Lck; and, 4) define the mechanisms that control LAT phosphorylation by ZAP-70.

尽管很可能已经确定了参与TCR信号的大多数分子，但我们对基础信号与诱导信号态的理解是基本的。酪氨酸激酶和磷酸酶保持动态平衡，该平衡控制和维持基础状态和其功能活性或空间定位的变化对于诱导的酪氨酸磷酸化很重要。该项目的重点是了解这些酪氨酸激酶和磷酸酶在质膜上的特异性和复杂调节。了解TCR细胞质Z链以及LAT蛋白的磷酸化的调节将是我们将重点关注的终点。我们将使用：纯化的重组蛋白；新型的遗传控制激酶抑制剂；二维脂质双层系统，其中包含定义的重组蛋白的定义量；并且，计算建模以描述和比较简单的双层系统与模型蜂窝系统的现有完整的完整CEKK或膜。我们将以迭代方式将这种二维模型的生化行为与蜂窝系统进行比较，从而提高组件复杂性并应用计算建模。我们期望这样的分析揭示预期和意外行为。我们还期望将简单系统与更复杂的系统进行比较，以揭示以前可能未被理解的调节途径的存在，例如正反馈和负反馈电路。我们清楚地认识到，我们无法对TCR信号的所有复杂性进行建模，但我们旨在了解控制TCR Z链的酪氨酸磷酸化的详细机制以及这些事件如何导致LAT磷酸化。项目＃1的总体目标是了解蛋白质酪氨酸激酶的特异性和调节参与TCR信号的磷酸酶在膜表面导致ITAM和LAT磷酸化。我们将：1）分别定义ITAM和LAT的LCK和ZAP-70的特异性分子基础； 2）为LCK开发一个模拟敏感抑制剂系统； 3）定义LCK控制ITAM磷酸化的机制； 4）定义通过ZAP-70控制LAT磷酸化的机制。