Evolution of proximal kinase network in T cells

T细胞中近端激酶网络的进化

基本信息

批准号：
10428138
负责人：
JOHN KURIYAN
金额：
$ 41.1万
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：
2011
资助国家：
美国
起止时间：
2011-07-15 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10428138
关键词：
Adaptive Immune System Adaptor Signaling Protein Affect Agonist Amino Acid Sequence Antigens B-Lymphocytes Behavior Binding Biochemical Biological Biophysics CD8B1 gene Cell Line Cell physiology Cells Charge Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Complex Diabetes Mellitus Dimerization Discrimination Engineering Environment Evolution Exhibits Family Family member Future Genes Genetic Transcription Glutamine Goals Hematopoietic Inbred NOD Mice Insulin-Dependent Diabetes Mellitus Jurkat Cells Knock-out LCP2 gene Ligands Lymphocyte-Specific p56LCK Tyrosine Protein Kinase Malignant Neoplasms Mediating Membrane Methods Molds Molecular Conformation Monitor Mus Mutagenesis Mutation Outcome PTPRC gene Peptide/MHC Complex Pharmaceutical Preparations Phosphatidylinositols Phosphoric Monoester Hydrolases Phosphorylation Phosphotransferases Phylogenetic Analysis Physiological Property Protein Dephosphorylation Protein Dynamics Protein Tyrosine Kinase Proteins Rest Role Scaffolding Protein Signal Pathway Signal Transduction Signal Transduction Pathway Signaling Molecule Signaling Protein Site Sorting - Cell Movement Stimulus Structure Substrate Specificity Syndrome T Cell Receptor Signaling Pathway T-Cell Development T-Cell Receptor T-Lymphocyte TEC Protein Tyrosine Kinase Tail Testing Trees Up-Regulation Variant ZAP-70 Gene base design exome sequencing experimental study fitness growth factor receptor-bound protein 2 inhibitor insight member membrane reconstitution mutant novel pathogen pressure programs receptor reconstitution reconstruction resistance mutation response single molecule src Homology Region 2 Domain src-Family Kinases

项目摘要

PROJECT 1 - ABSTRACT T cells are key components of the adaptive immune system, and have evolved to detect foreign antigens and generate a response that protects the host from intracellular pathogens and malignancies. The activation of the T cell receptor (TCR) results in the initiation of signal transduction pathways inside the T cell that generate the appropriate antigen-triggered responses. The major goals of the Program Project are to understand how the distinct features of the molecules and cellular circuitry in T cells allows for tonic signaling to self-pMHC, while also establishing a stimulus threshold, which when overcome results in robust signaling to agonists. In Project 1 we focus on the tyrosine kinases activated by the TCR, to understand the distinct properties of these signaling molecules, differentiating them from similar proteins operative in B cells. Our strategy is to use focused biochemical, biophysical, and cell biological studies on kinase variants and particular functions, combined with high-throughput methods for determining the fitness of variant proteins in supporting signal-transduction functions in T cells, as contrasted to B cells. The activation of intracellular signaling pathways by the TCR is mediated by three kinds of tyrosine kinases, which are members of the Src, Syk, and Tec families. Although they share features of their signaling machinery with other cells of the hematopoietic lineage, TCRs utilize a distinct set of members of these tyrosine kinase families (i.e., Lck, ZAP-70 and Itk), interact with unique MHC binding co-receptors (i.e., CD4 and CD8 with Lck) and phosphorylate unique scaffold proteins (e.g., LAT and SLP-76 for ZAP-70 and Itk) that couple to downstream signaling pathways. These components evolved contemporaneously with the MHC genes, and have maintained features in their sequences that mark them as distinct throughout the vertebrate evolutionary tree. A principal goal of Project 1 is to understand the functional specializations that have optimized these kinases for their roles in T cells. Project 1 has three Specific Aims. In Aim 1, we will define the specialized properties of the Src-family kinase Lck that are optimized for T cell function. These studies will include single-molecule tracking of Lck and variants to monitor their activation and localization, as well as structural studies on the interaction between Lck and the phosphatase CD45. In Aim 2 we will study the differentiation of ZAP-70 and Syk in T cells and B cells, by focusing on aspects of the activation of these kinases that differ in B cells and T cells. In Aim 3 we study the specialized differences in the activation mechanisms of Tec kinases, leading to different behavior on the membrane of Itk and Btk, which are the Tec kinases operative in T cells and B cells, respectively. Together, these studies will illuminate evolutionary pressures that have molded specialized responses from Lck, ZAP-70 and Itk, providing a framework for the development of T cell-specific drugs.

项目 1 - 摘要 T 细胞是适应性免疫系统的关键组成部分，并且已经进化到能够检测外来抗原和产生保护宿主免受细胞内病原体和恶性肿瘤侵害的反应。的激活 T 细胞受体 (TCR) 导致 T 细胞内信号转导途径的启动，从而产生适当的抗原触发反应。该计划项目的主要目标是了解如何 T 细胞中分子和细胞电路的独特特征允许向自身 pMHC 发出强直信号，同时还建立了一个刺激阈值，当克服该阈值时，会向激动剂发出强有力的信号。项目中 1 我们重点关注 TCR 激活的酪氨酸激酶，以了解这些信号传导的独特特性分子，将它们与 B 细胞中起作用的类似蛋白质区分开来。我们的策略是使用集中关于激酶变体和特定功能的生物化学、生物物理和细胞生物学研究，结合用于确定变异蛋白在支持信号转导中的适应性的高通量方法与 B 细胞相反，T 细胞发挥功能。 TCR 激活细胞内信号通路是由三种酪氨酸激酶介导的，它们是 Src、Syk 和 Tec 系列的成员。尽管它们具有相同的信号机制特征与造血谱系的其他细胞一样，TCR 利用这些酪氨酸激酶的一组不同成员家族（即 Lck、ZAP-70 和 Itk）与独特的 MHC 结合共受体（即 CD4 和 CD8 与 Lck）相互作用并磷酸化与下游偶联的独特支架蛋白（例如 ZAP-70 和 Itk 的 LAT 和 SLP-76）信号通路。这些成分与 MHC 基因同时进化，并保持它们序列中的特征使它们在整个脊椎动物进化树中具有独特的特征。一位校长项目 1 的目标是了解优化这些激酶的功能的专门化在T细胞中。项目 1 有三个具体目标。在目标 1 中，我们将定义 Src 家族激酶 Lck 的特殊特性针对 T 细胞功能进行了优化。这些研究将包括 Lck 及其变体的单分子追踪监测它们的激活和定位，以及 Lck 和磷酸酶CD45。在目标 2 中，我们将研究 ZAP-70 和 Syk 在 T 细胞和 B 细胞中的分化，重点关注这些激酶在 B 细胞和 T 细胞中的激活方面有所不同。在目标 3 中，我们研究专门的 Tec 激酶激活机制的差异，导致 Itk 膜上的不同行为和 Btk，分别是在 T 细胞和 B 细胞中起作用的 Tec 激酶。这些研究共同将阐明了形成 Lck、ZAP-70 和 Itk 专门反应的进化压力，提供开发 T 细胞特异性药物的框架。