REGULATION OF FOCAL ADHESION STRUCTURE

这项工作的长期目标是了解细胞如何粘附到 细胞外基质受到调节。 我们将研究蛋白质 粘着斑中的相互作用，这是细胞 粘附在细胞外基质上， 到膜上。 细胞粘附在胚胎发育过程中起着重要作用，当 基于信号刺激细胞迁移或粘附 从细胞外基质传递。 细胞粘附也是一个主要的 人类疾病的因素：血液中粘附调节不当 血小板可以产生动脉血凝块，导致中风和心脏病 攻击，以及通常发生在癌细胞中的粘附力丧失 有助于肿瘤转移。 细胞粘附涉及三组蛋白质： 形成细胞外基质的细胞， 细胞外基质（整合素）和细胞骨架蛋白 将细胞骨架锚在整合素上。 该项目的重点是 整合素及其与细胞骨架的连接。 本提案的具体目的是详细描述 α和β亚基的胞质尾部之间的相互作用 与细胞骨架的整合。 合成肽将用于 代表整联蛋白胞质尾。 这些肽将用于 在共沉淀测定、固相结合测定和亲和测定中， 色谱实验，以确定结合的细胞骨架蛋白 整合素胞质结构域。 从这些获得的数据 实验将被用来生成代表潜在的模型， 整合素和细胞骨架之间的联系。 这些模型将在 使用免疫沉淀技术在活细胞中进行测试， 显微注射

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.