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INTEGRIN ASSOCIATED PROTEIN/CD47 IS A THROMBOSPONDIN RECEPTOR

整合素相关蛋白/CD47 是一种血小板反应蛋白受体

基本信息

批准号：
8100483
负责人：
WILLIAM A FRAZIER
金额：
$ 38万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8100483
关键词：
Address Affect Affinity Avidity Binding Binding Sites Biological Assay Blood Platelets Blood Vessels Bone Marrow Bone Resorption C-terminal CD36 Antigens CD36 gene CD47 Antigen CD47 gene Cardiovascular Diseases Cardiovascular system Cells Cholesterol Complex Crystallization Cultured Cells Cyclic GMP Electron Microscopy Elements Fostering Grant Health Homologous Gene Hyperplasia Injury Integrin Binding Integrins Ischemia Knockout Mice Lateral Leukocytes Ligands Membrane Methods Modeling Molecular Monitor Mus Mutagenesis Mutation Necrosis Nitric Oxide Osteoclasts Oxygen Peptides Perfusion Phenotype Physiological Platelet Activation Play Process Property Proteins Regulation Resistance Role Signal Pathway Signal Transduction Site Smooth Muscle Myocytes Structure Sum Surface Testing Thrombospondin 1 Time Tissues Transgenes Vascular System X ray diffraction analysis X-Ray Crystallography X-Ray Diffraction angiogenesis base cell growth regulation design extracellular in vitro testing in vivo inhibitor/antagonist mutant novel novel strategies physical model receptor vasoconstriction

项目摘要

DESCRIPTION (provided by applicant): Thrombospondin-1 (TSP1) and its receptors have long been thought to have important roles in regulating vascular cells, both circulating and mural. During the preceding grant period we have discovered that TSP1 and CD47 (integrin-associated protein) regulate the dynamic range of nitric oxide (NO) signaling in vascular cells. Thus TSP1-CD47 interactions are important not only in angiogenic regulation but in rapid regulation of tissue perfusion and many other roles where NO maintains the health of the cardiovascular system. We have identified the binding surface on the C-terminal domain of TSP1 that interacts with CD47. We have also discovered a new extracellular mechanism for integrin activation via CD47. This proposal focuses on the molecular interactions among TSP1, CD47 and 23 integrins. The molecular details of these interactions will be deduced by mutagenesis of all three interacting partners. Mutant proteins will be tested in vitro in binding assays and assays of CD47 and integrin function. The extracellular clasp mechanism for integrin activation that we identified within the 1v23 structure will be tested in cultured cells and in 23-null mice repopulated with bone marrow expressing activated 23 integrin constructs. A similar approach will test the ability of CD47 to activate 23 integrins via interaction with the clasp in cultured cells and in mice expressing mutant CD47 transgenes. Conditions for formation of TSP1-CD47 and CD47-integrin complexes will be evaluated using physical methods and several EM approaches. These methods will then be used to evaluate the interactions of mutant proteins in order to identify structural elements of each that are important for their molecular interactions. Based on these results, crystallization and X-ray diffraction studies will be initiated of CD47 in complexes with the TSP1 C-terminal domain and with 1v23 integrin. The sum of these studies will provide for the first time a physical model of TSP1-CD47-integrin interactions. Given our new paradigm for TSP1-CD47 regulation of NO signaling in the vascular system, this information will be vital in designing new strategies to modulate endogenous NO signaling, a new approach to ameliorating cardiovascular disease.

描述（由申请人提供）：长期以来，血小板反应蛋白-1（TSP1）及其受体一直被认为在调节血管细胞（循环细胞和壁细胞）中具有重要作用。在之前的资助期间，我们发现 TSP1 和 CD47（整合素相关蛋白）调节血管细胞中一氧化氮 (NO) 信号的动态范围。因此，TSP1-CD47 相互作用不仅在血管生成调节中很重要，而且在组织灌注的快速调节以及 NO 维持心血管系统健康的许多其他作用中都很重要。我们已经鉴定了 TSP1 C 端结构域上与 CD47 相互作用的结合表面。我们还发现了一种通过 CD47 激活整合素的新细胞外机制。该提案重点关注 TSP1、CD47 和 23 整合素之间的分子相互作用。这些相互作用的分子细节将通过所有三个相互作用伙伴的诱变来推断。突变蛋白将在体外结合测定以及 CD47 和整合素功能测定中进行测试。我们在 1v23 结构中确定的整合素激活的细胞外扣机制将在培养细胞和用表达活化的 23 整合素构建体的骨髓重新填充的 23 缺失小鼠中进行测试。类似的方法将测试 CD47 通过与培养细胞和表达突变 CD47 转基因的小鼠中的扣环相互作用来激活 23 个整合素的能力。 TSP1-CD47 和 CD47-整合素复合物的形成条件将使用物理方法和几种 EM 方法进行评估。然后，这些方法将用于评估突变蛋白的相互作用，以确定每个突变蛋白对其分子相互作用重要的结构元件。基于这些结果，将启动 CD47 与 TSP1 C 末端结构域和 1v23 整联蛋白复合物的结晶和 X 射线衍射研究。这些研究的总和将首次提供 TSP1-CD47-整合素相互作用的物理模型。鉴于我们的 TSP1-CD47 调节血管系统中 NO 信号传导的新范例，这些信息对于设计调节内源性 NO 信号传导的新策略（一种改善心血管疾病的新方法）至关重要。