HUMAN ERYTHROCYTE MEMBRANE CYTOSKELETON ASSOCIATIONS

该提案的目标是 a) 更详细地阐明 人红细胞膜蛋白质的分子间缔合 骨架； b) 确定膜蛋白在红细胞生成过程中如何组装 发展； c) 使用红细胞膜作为实验 用于理解膜结构和功能的可访问模型系统 在更复杂的细胞中。 另一个令人感兴趣的新领域是 恶性疟原虫侵袭人红细胞的机制 寄生虫。 作为一般方法，这些研究将通过 已纯化和表征的膜蛋白，以及 这些蛋白质之间的关联将被定量评估。 具体项目包括以下几个方面： 1) 结合的研究 红细胞锚蛋白和锚蛋白的蛋白水解片段在溶液中 确定的血影蛋白寡聚状态和条带的细胞质结构域 3. 2) 对新发现的钙调蛋白结合蛋白的研究 红细胞膜骨架的组成部分，其目标是确定 它的蛋白质关联，以及钙对这些关联的调节 和钙调蛋白。 3) 将 RBC 作为研究模型系统进行评估 受体介导的内吞作用中的蛋白质关联，使用网格蛋白 最近已从红细胞胞浆中纯化出转铁蛋白受体 胎盘。 4) 迄今为止尚未表征的红细胞蛋白的研究 包括纯化、最近邻分析和异构体搜索 在其他组织中。 候选者将是带 4.2，一种膜骨架形式 带 7 和最近发现的钙调蛋白结合蛋白。 5) 组装 红白血病分化过程中红细胞膜骨架的变化 细胞。 6) 人红细胞侵袭机制的研究 恶性疟原虫裂殖子，重点是裂殖子的纯化 与带 3 相互作用的蛋白质。

The goals of this proposal are to a) elucidate in further detail the intermolecular associations of proteins of the human erythrocyte membrane skeleton; b) determine how membrane proteins assemble during erythroid development; c) use the erythrocyte membrane as an experimentally accessible model system for understanding membrane structure and function in more complex cells. An additional, new area of interest is the mechanism of invasion of human erythrocytes by P. falciparum malarial parasites. As a general approach, these studies will be performed with membrane proteins that have been purified and characterized, and associations between these proteins will be evaluated quantitatively. Specific projects are in the following areas: 1) Studies of binding of erythrocyte anykrin and proteolytic fragments of ankyrin in solution to defined oligomeric states of spectrin and the cytoplasmic domain of band 3. 2) Studies of a newly discovered calmodulin-binding protein that is a component of the erythrocyte membrane skeleton with goals of determining its protein associations, and regulation of these associations by calcium and calmodulin. 3) Evaluation of the RBC as a model system to study protein associations in receptor-mediated endocytosis, using clathrin that has recently been purified from RBC cytosol, and transferrin receptor from placenta. 4) Studies of heretofore uncharacterized erythrocyte proteins including purification, nearest neighbor analysis and search for isoforms in other tissues. Candidates will be band 4.2, a membrane skeletal form of band 7 and a recently discovered calmodulin-binding protein. 5) Assembly of an erythroid membrane skeleton during differentiation of erythroleukemia cells. 6) Studies of the mechanisms of invasion of human erythrocytes by merozoites of P. falciparum, with emphasis on purification of merozoite proteins that interact with band 3.