PLATELET ACTIVATION AND MICROFILAMENT ORGANIZATION

本研究的目的是了解 黏着斑蛋白在血小板活化和微丝组织中的作用。 我们 工作假设是黏着斑蛋白以几种形式存在， 血小板活化这些形式中的一种或多种被修饰以允许其 与肌动蛋白、其他肌动蛋白相关蛋白或血浆的相互作用 膜的 为了验证这一假设，我们计划分离细胞骨架和 来自未活化和活化血小板的膜部分和用于 免疫印迹分析黏着斑蛋白的分布和定位。 我们也 将研究可溶性血小板黏着斑蛋白的体外相互作用， 用粘度测定法纯化了肌动蛋白和肌动蛋白结合蛋白， 荧光、沉降、亲和基质和凝胶覆盖分析。 此外，我们将研究钙和磷酸化对细胞增殖的影响。 黏着斑蛋白在未活化的、活化的、粘附的、 和聚集的血小板。 这些研究提供了基本的 黏着斑蛋白在细胞骨架组织中的作用， 血小板活化

The objective of this research proposal is to understand the role of vinculin in platelet activation and microfilament organization. Our working hypothesis is that vinculin exist in several forms and following platelet activation one or more of these forms is modified to allow its interaction with actin, other actin associated proteins, or the plasma membrane. To test this hypothesis we plan to isolate cytoskeletal and membrane fractions from unactivated and activated platelets and probe for vinculin's distribution and localization by immunoblot analysis. We also will study the in vitro interaction of the soluble platelet vinculin we have purified with actin and actin binding proteins using viscometric, fluorometric, sedimentation, affinity matrix and gel overlay analyses. Further, we will study the effects of calcium and phosphorylation on the distribution and activity of vinculin in unactivated, activated, adherent, and aggregated platelets. These studies should provide fundamental information on the role of vinculin in cytoskeletal organization and also in platelet activation.