RED CELL MEMBRANE SKELETON; ULTRASTRUCTURE & FUNCTION

红细胞膜骨架；

• 批准号: 3344931
• 负责人: BETTY W SHEN
• 金额: $ 14.2万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3344931
• 关键词: actins; blood viscosity; cell age; cell biology; chemical structure function; chromatography; conformation; congenital hemolytic anemia; cryoscience; cytoskeleton; electron microscopy; electrophoresis; erythrocyte membrane; erythrocytes; human tissue; immunochemistry; laboratory mouse; laboratory rabbit; membrane proteins; membrane structure; microfilaments; morphology; reticulocytes; sickle cell anemia; spectrin; tropomyosin

We propose to continue our analysis of the fine structures of the network that lines the cytoplasmic surface of the human red cell membrane and to analyze the structural changes occurring to this skeletal network during the maturation of reticulocytes. The goal of this study is a better understanding of both the molecular architecture of the membrane skeleton and the fundamental basis for the durability, deformability, and elasticity of the normal red cell and the precise nature of the structural derangements in the skeletons of pathological red cells. We have shown by electron microscopic examination of negatively stained specimens that the skeleton is organized into short actin protofilaments interconnected by multiple strands of spectrin tetramer. The spectrin, which is the major element of the membrane skeleton, is in a compact or condensed form when freshly released from the static membranes but can be extended by both electrostatic and mechanical forces to the elongated and filamentous form commony observed for isolated spectrin. We propose to extend our investigation of the red cell membrane skeleton by examinung the following factors: (1) The conformation of spectrin and its contribution to the elasticity of the skeleton, (2) The basic organization of membrane skeleton of the human erythrocytes, and (3) The role of the skeleton in maintaining the shape and mechanical properties of the red cells. A variety of experimental approaches will be used including immunofluorescence microscopy, immunoelectron microscopy, and low-dose cryoelectronmicroscopy of frozen-hydrated specimens, as well as other biochemical and biophysical techniques. The principal aims will be to elucidate the precise nature and cellular significance of skeletal elasticity. Information gained from the above studies will help elucidate the nature of the structural derangement in the skeletons of pathological red cells and may also assist in the analysis of cytoskeletons of nucleated cells.

我们建议继续对精细结构进行分析 排列在人类红细胞细胞质表面的网络 膜并分析其发生的结构变化 网织红细胞成熟过程中的骨骼网络。 目标 这项研究的目的是更好地了解分子 膜骨架的结构和基本原理 具有普通红色的耐用性、可变形性和弹性 细胞和结构紊乱的精确性质 病理性红细胞的骨架。 我们通过电子显微镜检查显示出阴性 染色标本表明骨骼被组织成短肌动蛋白 由多股血影蛋白相互连接的原丝 四聚体。 血影蛋白，它是血影蛋白的主要元素 膜骨架，当处于致密或凝聚形式时 刚从静态膜中释放出来，但可以延长 通过静电力和机械力来拉长和 分离的血影蛋白常见丝状形式。 我们 提议扩大我们对红细胞膜的研究 通过检查以下因素来确定骨架： (1) 血影蛋白的构象及其对弹性的贡献 (2)膜骨架的基本组织 人类红细胞，以及（3）骨骼在 保持红细胞的形状和机械特性。 将使用各种实验方法，包括 免疫荧光显微镜、免疫电子显微镜、 以及冷冻水合物的低剂量冷冻电子显微镜 标本以及其他生物化学和生物物理 技术。 主要目标是阐明准确的 骨骼弹性的性质和细胞意义。 从上述研究中获得的信息将有助于阐明 骨骼结构紊乱的本质 病理性红细胞，也可能有助于分析 有核细胞的细胞骨架。