REGULATION OF FOCAL ADHESION STRUCTURE

这项工作的长期目标是了解细胞如何黏附到 细胞外基质受到调节。我们将研究这种蛋白质 局灶性粘连中的相互作用，这是细胞 附着在细胞外基质和细胞骨架锚定的地方 到膜上。 细胞黏附在胚胎发育过程中起着重要作用，当 根据信号刺激细胞迁移或黏附 从细胞外基质输送过来的。细胞粘附性也是一个主要的 人类疾病中的因素：血液黏附调节不当 血小板可产生动脉血栓，导致中风和心脏 攻击，以及通常发生在癌细胞中的粘附性丧失 会导致肿瘤转移。 细胞黏附涉及三组蛋白质：细胞外的蛋白质 形成细胞外基质的细胞，跨膜蛋白结合 细胞外基质(整合素)和细胞骨架蛋白 将细胞骨架固定在整合素上。这个项目是专注于 关于整合素及其与细胞骨架的联系。 这项提案的具体目的是详细说明 α和β亚基胞质尾部之间的相互作用 与细胞骨架的整合。合成肽将被用来 代表整合素胞质尾巴。这些多肽将被用于 在共沉淀分析、固相结合分析和亲和分析中 鉴定结合细胞骨架蛋白的层析实验 整合素胞浆域。从这些数据中获得的数据 实验将被用来生成代表潜力的模型 整合素和细胞骨架之间的联系。这些模型将是 在活细胞中使用免疫沉淀和 微量注射。

The long-term goal of this work is to understand how cell adhesion to the extracellular matrix is regulated. We will study the protein interactions in the focal adhesions, which are the sites where cells adhere to the extracellular matrix and where the cytoskeleton is anchored to the membrane. Cell adhesion plays an important role during embryonic development, when cells are stimulated either to migrate or adhere based on signals conveyed from the extracellular matrix. Cell adhesion is also a major factor in human disease: improper regulation of adhesion in blood platelets can generate arterial blood clots that cause strokes and heart attacks, and the loss of adhesion that typically occurs in cancer cells contributes to tumor metastasis. Cell adhesions involves three groups of proteins: proteins outside the cell that form the extracellular matrix, transmembrane proteins that bind to the extracellular matrix (the integrins), and cytoskeleton proteins that anchor the cytoskeleton to the integrins. This project is focused on the integrins and their linkage to the cytoskeleton. The specific aims of this proposal are to characterize in detail the interactions between the cytoplasmic tails of the alpha and beta subunits of intergrin with the cytoskeleton. Synthetic peptides will be used to represent the integrin cytoplasmic tails. These peptides will be used in co-precipitation assays, solid-phase binding assays, and affinity chromatography experiments to identify cytoskeletal proteins that bind to the integrin cytoplasmic domains. The data obtained from these experiments will be used to generate models representing potential linkages between integrins and the cytoskeleton. These models will be tested in living cell using the techniques of immunoprecipitation and microinjection.