Deconvoluting the Vascular Adhesome

去卷积血管粘附体

• 批准号: 10327637
• 负责人: Mark HOWARD Ginsberg
• 金额: $ 78.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10327637
• 关键词: Accounting; Adhesions; Binding Sites; Biochemical; Blood; Blood Platelets; Blood Vessels; Cell Adhesion; Cell Line; Cells; Complex; Data; Endothelial Cells; Family; Foundations; Future; Health behavior; Heterogeneity; Image; Integrin alpha Chains; Integrin beta Chains; Integrins; Leukocytes; Ligation; Mesenchymal; Methods; Molecular; Myocardial Infarction; Output; Process; Proteins; Regulation; Resources; Rod; Signal Transduction; Site; Stroke; Structure; Talin; Testing; Vinculin; base; cell motility; density; dynamical evolution; experimental study; member; novel strategies; physical state; therapeutic target

Summary Integrin-based adhesions are central to the functions of blood and vascular cells. The heterogeneity of adhesions and their dynamic evolution has complicated efforts to study their fine composition, assembly, and disassembly. Our studies have shown that a transition between two adhesion archetypes is controlled by a simple binary molecular switch of vinculin competition with and displacement of RIAM or lamellipodin(Lpd), members of the MRL family, from binding sites on talin's rod domain. Furthermore, we developed methods to image the complex of MRL proteins with integrins and talin (MIT complex) and showed that it formed the tips of “sticky fingers,” cellular protrusions that sense the density of matrix proteins and physical state of the substrate during mesenchymal cell migration. mesenchymal cell migration. We hypothesize that the MIT complex represents one among many distinct modules that contribute to the overall structure and function of integrin- based adhesions. This suggests the paradigm that the integrin adhesome can be analyzed as a dynamic assembly of these modules, which form prior to entry into the adhesions. This concept has enabled us to propose a new approach to studying adhesions by developing methods to purify each module formed prior to integrin ligation. To test this paradigm, we propose to purify 4 such modules and to characterize each for a) the presence of talin-activated integrins b) characterize its protein composition. c) establish its biochemical topology and regulation d) visualize it in living cells and e) evaluate its functions The proposed experiments will generate foundational data for the adhesion field in three ways: a) the integrin adhesome will be re- interpreted as a compendium of modular components; each with its own specific composition. b) The functional studies will identify new regulators of blood and vascular cell adhesion and signaling that may be exploitable as therapeutic targets c) A complete definition of the protein composition of each module will serve as a public resource for the analysis of integrin-based adhesions and will be hypothesis-generating for future studies to understand the regulation of assembly of each module and its functional outputs.

概括 基于整联蛋白的粘连是血液和血管细胞功能的核心。这 广告的异质性及其动态演变已经复杂地研究了他们的罚款 组成，组装和拆卸。我们的研究表明，两个 粘附原型受Vinculin竞争的简单二进制分子开关控制 MRL家族成员Riam或Lamellipodin（LPD）的位移来自绑定位点 在塔林的杆域中。此外，我们开发了成像MRL蛋白复合物的方法 与整合素和塔林（MIT复合物）一起表明它形成了“粘手指”的尖端， 感知基质蛋白的密度和底物的物理状态的突出 间充质细胞迁移。间充质细胞迁移。我们假设麻省理工学院 复合体是许多有助于整体的不同模块之一 基于整合素的广告的结构和功能。这表明范式 可以将整联蛋白拟合分析为这些模块的动态组装，在此之前形式 进入粘连。这个概念使我们能够提出一种新的研究方法 通过开发在整合素结扎之前纯化的每个模块的方法来粘附。测试 这种范式，我们建议净化4个这样的模块，并为每个模块表征a）存在 塔林激活的整联蛋白b）表征其蛋白质成分。 c）建立生化 拓扑和调节d）在活细胞中可视化它，e）评估其功能 实验将以三种方式生成粘附场的基础数据：a）整合素 伪造者将被解释为模块化成分的汇编；每个人都有自己的 特定组成。 b）功能研究将确定血液和血管的新调节剂 细胞粘附和可能作为治疗靶标的可以利用的信号传导c）一个完整的定义 每个模块的蛋白质成分将作为分析的公共资源 基于整合素的广告，并将为未来的研究提供假设，以了解 调节每个模块的组装及其功能输出。