MEMBRANE DYNAMICS IN NORMAL AND ABNORMAL RED BLOOD CELLS

正常和异常红细胞的膜动力学

• 批准号: 3344389
• 负责人: DAVID E. GOLAN
• 金额: $ 34.91万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1993-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3344389
• 关键词: band 3 protein; blood fractionation; calcium; calmodulin; congenital blood disorder; cytoskeleton; erythrocyte membrane; fluorescence; fluorescence microscopy; glycophorin; glycoproteins; hemolytic anemia; human tissue; membrane activity; membrane proteins; molecular dynamics; nuclear magnetic resonance spectroscopy; reticulocytes

The human red blood cell membrane is a dynamic structure which must respond to changes in its environment in order to assure continued cell viability. The submenbranous erythrocyte cytoskeleton plays a crucial role in defining red cell stability, deformability, and shape, properties which, in turn, influence interactions between the red cell and cells of the reticuloendothelial system. This proposal focuses on a set of dynamic membrane phenomena which are controlled by the cytoskeleton: the distribution and lateral mobility of the major transmembrane proteins of the human red blood cell -- band 3 and glycophorin. These phenomena will be examined on membranes which have been labeled with covalent fluorescent reagents specific for band 3 and/or glycophorin, using the techniques of fluorescence microscopy, fluorescence phtobleaching recovery, and fluorescence energy transfer. The intermolecular interactions among band 3, glycophorin, and cytoskeletal components will be probed, differentiating among various dynamic control mechanisms such as steric hindrance, specific binding, and complex formation. The mechanism of immobilization of glycophorin in the membrane by elevated intracellular calcium, and the possible role of calmodulin therein, will also be investigated. Detailed study of transmembrane protein dynamics in normal and abnormal erythrocyte membranes -- such as those from patients with hereditary spherocytosis, hereditary elliptocytosis, hereditary pyropoikilocytosis and sickle cell anemia -- is expected to provide new information about the structure and function of the red cell membrane in health and disease. Alterations in transmembrane protein dynamics may, for example, adversely affect interactions between red cells and cells of the reticuloendothelial system, leading to abnormal adherence, sequenstration, or hemolysis. The goal of this research is the understanding, in molecular terms, of the cytoskeletal forces controlllng the distribution and lateral mobility of transmembrane proteins in the human erythrocyte membrane, and of the relationship between derangements in these control mechanisms and the pathophysiology of disease in various abnormal red cell membrane states.

人类红细胞膜是一种动态结构，必须响应 为了确保持续的细胞生存能力。 下属红细胞细胞骨架在定义中起着至关重要的作用 红细胞的稳定性，可变形性和形状，属性又 影响红细胞与细胞之间的相互作用 网状内皮系统。 该提案着重于一组动态 由细胞骨架控制的膜现象： 主要跨膜蛋白的分布和横向迁移率 人类红细胞-Band 3和Glyocophorin。 这些现象会 在用共价荧光标记的膜上检查 对条带3和/或糖果蛋白特异的试剂，使用的技术 荧光显微镜，荧光phtoblesing恢复和 荧光能量转移。 带之间的分子间相互作用 将探测3，糖脂蛋白和细胞骨架成分，并区分 在各种动态控制机制（例如空间障碍）中， 结合和复杂形成。 固定的机制 通过升高的细胞内钙在膜中的糖蛋白，并 其中的钙调蛋白的可能作用也将被研究。 详细的 跨膜蛋白动力学在正常和异常红细胞中的研究 膜 - 例如来自遗传性球细胞增多症患者的膜， 遗传性椭圆细胞增多症，遗传性心形细胞增多症和镰状细胞 贫血 - 有望提供有关结构和 红细胞膜在健康和疾病中的功能。 改变 跨膜蛋白动力学可能会对 红细胞与网状内皮系统细胞之间的相互作用， 导致异常依从性，序列或溶血。 目标 这项研究是从分子角度对细胞骨架的理解 力控制跨膜的分布和横向迁移率 人类红细胞膜中的蛋白质以及 这些控制机制和疾病的病理生理学的危险 在各种异常红细胞膜状态下。