Evolution of proximal kinase network in T cells

T细胞中近端激酶网络的进化

• 批准号: 10615817
• 负责人: JOHN KURIYAN
• 金额: $ 40.07万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 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/10615817
• 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; Stimulus; Structure; Substrate Specificity; Syndrome; T Cell Receptor Signaling Pathway; T-Cell Activation; T-Cell Development; T-Cell Receptor; T-Lymphocyte; TEC Protein Tyrosine Kinase; Tail; Testing; Trees; Up-Regulation; Variant; ZAP-70 Gene; design; exome sequencing; experimental study; fitness; growth factor receptor-bound protein 2; inhibitor; inorganic phosphate; insight; member; membrane reconstitution; mutant; novel; nuclear factors of activated T-cells; pathogen; pressure; programs; protein purification; receptor; reconstitution; reconstruction; resistance mutation; response; single molecule; src Homology Region 2 Domain; src-Family Kinases

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细胞是自适应免疫系统的关键组成部分，并且已进化为检测外国抗原和 产生一种保护宿主免受细胞内病原体和恶性肿瘤的反应。激活 T细胞受体（TCR）导致启动T细胞内部信号转导途径，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 T细胞中分子和细胞电路的不同特征允许对自PMHC进行强直信号传导，而 还建立一个刺激阈值，当克服时，它会导致对激动剂的强大信号。在项目中 1我们专注于TCR激活的酪氨酸激酶，以了解这些信号的不同特性 分子，将它们与B细胞中相似蛋白的手术区分开。我们的策略是使用专注 关于激酶变异和特定功能的生化，生物物理和细胞生物学研究，并结合 用于确定变异蛋白在支持信号转导的适应性的高通量方法 与B细胞形成鲜明对比的T细胞的功能。 TCR的细胞内信号通路的激活是由三种酪氨酸激酶介导的 是SRC，SYK和TEC家庭的成员。尽管他们共享信号机械的功能 与造血谱系的其他细胞一起，TCR使用了这些酪氨酸激酶的一组不同的成员 家族（即LCK，ZAP-70和ITK）与唯一的MHC结合共受体（即与LCK的CD4和CD8）相互作用 和磷酸化的独特脚手架蛋白（例如，ZAP-70和ITK的LAT和SLP-76）夫妇到下游 信号通路。这些成分与MHC基因同时发展，并保持 其序列中的特征在整个脊椎动物进化树中标记为不同。校长 项目1的目标是了解为其角色优化这些激酶的功能专业 在T细胞中。 项目1具有三个特定目标。在AIM 1中，我们将定义SRC家庭激酶LCK的专业特性 针对T细胞功能进行了优化。这些研究将包括LCK和变体的单分子跟踪 监控其激活和定位，以及有关LCK与LCK之间相互作用的结构研究 磷酸酶CD45。在AIM 2中，我们将通过聚焦在T细胞和B细胞中ZAP-70和SYK的分化 在这些激活的各个方面，这些激酶在B细胞和T细胞中不同。在AIM 3中，我们研究专业 TEC激酶激活机制的差异，导致ITK膜上不同的行为 BTK和BTK分别是TEC激酶在T细胞和B细胞中的作战。这些研究将在一起 照明从LCK，ZAP-70和ITK塑造了专门响应的进化压力，提供了 T细胞特异性药物开发的框架。