HUMAN ERYTHROCYTE MEMBRANE CYTOSKELETON ASSOCIATIONS

本提案的目标是：a）进一步详细阐明 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系期间如何组装 c）使用红细胞膜作为实验 了解膜结构和功能的可访问模型系统 更复杂的细胞。 另一个新的关注领域是 恶性疟原虫侵入人红细胞的机制 寄生虫 作为一般方法，这些研究将采用 已经纯化和表征的膜蛋白，和 将定量评价这些蛋白质之间的关联。 具体项目包括以下几个方面：（1）研究 红细胞锚蛋白和锚蛋白的蛋白水解片段， 血影蛋白的确定的寡聚状态和条带的胞质结构域 3. 2)一种新发现的钙调素结合蛋白的研究 红细胞膜骨架的组成部分，目的是确定 它的蛋白质协会，并通过钙调节这些协会 和钙调素。 3)RBC作为研究模型系统的评价 蛋白质协会在受体介导的内吞作用，使用网格蛋白， 最近从RBC胞质溶胶中纯化，转铁蛋白受体从 胎盘 4)红细胞蛋白质的研究 包括纯化、最近邻分析和寻找同种型 在其他组织中。 候选者将是条带4.2，一种膜骨架形式， 带7和最近发现的钙调素结合蛋白。 5)组件 红系细胞膜骨架在红白血病分化过程中的变化 细胞 6)人红细胞侵袭机制的研究 恶性疟原虫裂殖子的分离纯化 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.