Regulation of Integrin Signaling by Adapter Proteins

接头蛋白对整合素信号传导的调节

• 批准号: 7323902
• 负责人: GARY A. KORETZKY
• 金额: $ 30.68万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7323902
• 关键词: Address; Adhesions; Alleles; Animals; Avidity; Binding; Biochemical; Biochemistry; Breeding; Cell Adhesion; Cell Lineage; Cell Surface Receptors; Cell physiology; Cell surface; Cells; Complement; Complex; Condition; Data; Distal; Dominant-Negative Mutation; Event; Family member; Fc Receptor; Gene Targeting; Genetic; Goals; Guanosine Triphosphate Phosphohydrolases; Hematopoietic; Image; Immune; Immune system; Integrin Binding; Integrin Signaling Pathway; Integrins; Knock-in Mouse; Laboratories; Lead; Learning; Leukocytes; Ligands; Localized; Location; Mature T-Lymphocyte; Membrane; Membrane Microdomains; Modeling; Molecular; Molecular Conformation; Molecular Genetics; Molecular Weight; Movement; Mus; Mutation; Numbers; Pathway interactions; Peripheral; Phosphoproteins; Play; Positioning Attribute; Property; Proteins; Proteomics; Recruitment Activity; Regulation; Research Personnel; Rest; Role; SH3 Domains; Second Messenger Systems; Series; Signal Pathway; Signal Transduction; Surface; T-Cell Development; T-Lymphocyte; Testing; Transgenes; Tyrosine Phosphorylation; Variant; Work; adapter protein; base; cell motility; cell type; design; in vivo; insight; interest; intermolecular interaction; intracellular protein transport; migration; mutant; neutrophil; programs; protein function; protein localization location; receptor; research study; response; second messenger; src Homology Region 2 Domain; src-Family Kinases; tool

Integrins are dimeric cell surface receptors that play critical roles in cellular adhesion and migration. Signaling events critical for integrin function include second messenger cascades initiated by other cell surface receptors which modulate integrin avidity for their ligands in addition to signaling pathways initiated by engagement of integrins themselves. We and others have found that the formation of multimolecular complexes nucleated by adapater proteins is essential for both signaling to integrins (inside-out signaling) and signaling by integrins (outside-in signaling). Our laboratory has a long standing interest in the SH2 domain containing leukocyte phosphoprotein of 76kDa (SLP-76), a hematopoietic restricted adapater protein that is critical for function of both immunoreceptors and integrins. The experiments described in this proposal will make use of biochemical, imaging, molecular and genetic approaches to investigate the role of SLP-76 and its associated molecules in integrin responses in cells of both the innate and adaptive immune systems. The work will be divided into three specific aims. The first will explore the inducible interactions between SLP-76 and other proteins upon integrin engagement and the subcellular localization of SLP-76 during integrin function in T cells. One central hypothesis that will be tested is that there are two pools of SLP-76 in the cell, one which regulates immunoreceptor signaling and the second which regulates integrin responses. We will further test the notion that integrin pathway requires an interaction between SLP-76 and three adapter proteins, ADAP (Adhesion and Degranulation-promoting Adapter Protein), SKAP55 (SrcKinase- associated phosphoprotein of 55kDa), and RIAM (Rap1-GTP interacting adapter molecule), and that these adapters collectively nucleate a complex which brings activated Rap-1 to the cell surface. The second aim of this proposal will take the biochemical and imaging studies of Aim 1 in vivo by generating genetically unique murine lines in which mutations of SLP-76 or its associated molecules are expressed that are predicted to selectively impact T cell immunoreceptor or integrin function. The third aim will extend our work to the innate immune system by studying the biochemistry and molecular events coordinated by these adapter molecules in neutrophils, both ex vivo and in vivo. Experiments described in this project will make extensive use of both scientific cores and will rely extensively on interactions with the Project Leaders of the other projects of this program. We hope that these studies will provide new insights into how multimolecular complexes integrate signaling pathways leading to appropriate immune cell responses.

整合素是在细胞粘附和迁移中起关键作用的二聚体细胞表面受体。 对整合素功能至关重要的信号事件包括由其他细胞启动的第二信使级联反应， 调节整合素对其配体的亲合力的表面受体以及启动的信号传导途径 通过整合素本身的参与。我们和其他人已经发现多分子的形成 由衔接子蛋白质成核的复合物对于向整合素的信号传导（由内向外信号传导）是必需的。 和通过整合素的信号传导（由外向内信号传导）。我们的实验室对SH 2有着长期的兴趣 含76 kDa白细胞磷蛋白（SLP-76）的结构域，一种造血限制性衔接蛋白 这对于免疫受体和整合素的功能都是至关重要的。本提案中所述的实验 将利用生物化学、成像、分子和遗传学方法来研究SLP-76的作用 及其相关分子在先天性和适应性免疫系统细胞中的整合素应答中的作用。 这项工作将分为三个具体目标。第一个将探索诱导的相互作用， SLP-76和其他蛋白质在整合素接合时的作用以及SLP-76在细胞内的亚细胞定位 整合素在T细胞中的功能。将被检验的一个中心假设是，存在两个SLP-76池， 细胞，一个调节免疫受体信号传导，第二个调节整合素反应。 我们将进一步测试整合素途径需要SLP-76和三个细胞因子之间相互作用的概念。 衔接蛋白，ADAP（粘附和脱粒促进衔接蛋白），SKAP 55（SrcKinase- 55 kDa的相关磷蛋白）和RIAM（Rap 1-GTP相互作用衔接分子），并且这些 衔接子共同使复合物成核，该复合物将活化的Rap-1带到细胞表面。第二个目的 该提案的一部分将通过遗传学方法对Aim 1进行体内生物化学和成像研究， 表达SLP-76或其相关分子突变的独特鼠系， 预测选择性地影响T细胞免疫受体或整合素功能。第三个目标将扩展我们的工作 先天免疫系统的生物化学和分子事件的研究协调这些 嗜中性粒细胞中的衔接子分子，包括离体和体内。本项目中描述的实验将使 广泛使用这两个科学核心，并将广泛依赖与项目负责人的互动， 这个项目的其他项目。我们希望这些研究将为多分子如何 复合物整合了导致适当免疫细胞应答的信号传导途径。