PILOT STUDY--TRANSMEMBRANE GLYCOPROTEIN INTERACTIONS

细胞骨架-跨膜糖蛋白相互作用具有 在各种细胞功能中发挥重要作用，包括细胞 生长、细胞黏附和信号转导。几种蛋白质 调节细胞骨架与蛋白质和蛋白质的相互作用 细胞膜上的结构。ERM蛋白(Ezrin、Moesin和 Radixin)是一个新发现的家族，被认为是膜- 细胞骨架连接物。基因中的突变和扰动 细胞骨架-膜连接蛋白，包括ERM蛋白，可以 干扰正常的细胞黏附。通过这种方式，细胞可以获得 致癌特性。 我们建议研究跨膜糖蛋白，如 细胞黏附分子C-CAM105，硫酸肝素 蛋白多糖Syndecan-1和上皮粘蛋白MUC1是 与细胞骨架锚定蛋白或其他ERM相关 蛋白质。跨膜糖蛋白的胞质尾巴 将用于亲和层析研究以检测 代谢标记细胞裂解产物中的相互作用蛋白。这个 已确定的相互作用将与纯化的蛋白质和 将对这些蛋白质上的结合部位进行表征。这个 Ezrin与跨膜糖蛋白的结合也将是 Ezrin基因导入培养细胞的实验研究 确定Ezrin是否重新分布跨膜 糖蛋白及其分子决定簇的测定 将阐明Ezrin或其他蛋白的作用 细胞骨架分子在C-CAM105、MUC1和 Syndecan-1。

Cytoskeletal-transmembrane glycoprotein interactions have an important role in various cellular functions, including cell growth, cell adhesion, and signal transduction. Several proteins mediate the interactions of the cytoskeleton with proteins and structures on the cell membrane. ERM proteins (ezrin, moesin and radixin) are a newly found family postulated to be membrane- cytoskeleton linkers. The mutation and perturbations in the cytoskeletal-membrane linker proteins, including ERM proteins, may disturb normal cell adhesion. In this way the cells could obtain tumorigenic properties. We propose to study whether transmembrane glycoproteins such as the cell adhesion molecule C-CAM 105, the heparin sulfate proteoglycan syndecan-1, and the epithelial mucin MUC1 are associated with cytoskeletal anchorage proteins or other ERM proteins. The cytoplasmic tails of transmembrane glycoproteins will be used in affinity chromatography studies to detect interacting protein in metabolically labeled cell lysates. The identified interactions will be studied with purified proteins and the binding sites on these proteins will be characterized. The association of ezrin with transmembrane glycoproteins will also be studied by transfection of ezrin cDNA into cultured cells to determine whether ezrin redistributes the transmembrane glycoproteins and the determination of the molecular determinants of protein interactions will clarify the role of ezrin or other cytoskeletal molecules in the function of C-CAM 105, MUC1, and syndecan-1.