PLATELET ACTIVATION AND MICROFILAMENT ORGANIZATION

这项研究提案的目的是了解 纽蛋白参与血小板活化和微丝组织。我们的 工作假说是纽蛋白以几种形式存在 这些形式中的一种或多种被修改以允许其 与肌动蛋白、其他肌动蛋白相关蛋白或血浆的相互作用 薄膜。为了验证这一假设，我们计划分离细胞骨架和 来自未激活和激活的血小板的膜组分和探针 免疫印迹法检测纽蛋白的分布和定位。我们也 将研究可溶性血小板纽蛋白WE的体外相互作用 已经用肌动蛋白和肌动蛋白结合蛋白进行了纯化， 荧光法、沉降法、亲和力矩阵和凝胶覆盖分析。 进一步，我们将研究钙和磷酸化对血管内皮细胞的影响。 纽蛋白在未活化、活化、贴壁、 和聚集的血小板。这些研究应该提供基本的 纽蛋白在细胞骨架组织中的作用以及 在血小板活化方面。

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.