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细胞是适应性免疫系统的关键组成部分，它已经进化到能够检测外来抗原和 产生一种反应，保护宿主免受细胞内病原体和恶性肿瘤的侵袭。激活 T细胞受体(TCR)导致启动T细胞内的信号转导通路，从而产生 适当的抗原触发的反应。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向自身pMHC发出紧张性信号，而 还建立了一个刺激阈值，当超过这个阈值时，会向激动剂发出强有力的信号。在项目中 1我们重点研究由TCR激活的酪氨酸激酶，以了解这些信号的不同属性。 分子，将它们与在B细胞中起作用的类似蛋白质区分开来。我们的策略是使用专注的 生物化学、生物物理学和细胞生物学研究，结合 确定支持信号转导的不同蛋白质适合性的高通量方法 T细胞的功能，而不是B细胞。 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家族的成员。尽管它们具有相同的信令机制特征 与造血系的其他细胞一起，TCR利用这些酪氨酸激酶的一组不同的成员 家族(即Lck、ZAP-70和ITK)与独特的MHC结合共受体(即与Lck结合的CD4和CD8)相互作用 并磷酸化连接到下游的独特支架蛋白(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，并保持了 在整个脊椎动物进化树中，将它们标记为不同的特征。一位校长 项目1的目标是了解针对其角色优化这些Kase的功能专业化 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的特殊性质 它们是为T细胞功能而优化的。这些研究将包括对LCK及其变异体的单分子跟踪 监测它们的激活和定位，以及对Lck和Lck之间相互作用的结构研究 磷酸酶CD45。在目标2中，我们将通过聚焦研究ZAP-70和Syk在T细胞和B细胞中的分化 在这些不同于B细胞和T细胞的激酶的激活方面。在目标3中，我们学习专门的 TEC激酶激活机制的不同，导致ITK膜上的行为不同 和BTK，它们分别是在T细胞和B细胞中工作的Tec激酶。总而言之，这些研究将 阐明已经塑造了LCK、ZAP-70和ITK的专门反应的进化压力，提供 开发T细胞特异性药物的框架。