CELLULAR & MOLECULAR BIOLOGY OF ALTERED BAND 3

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• 批准号: 3120302
• 负责人: MARGUERITE M KAY
• 金额: $ 7.24万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-10-01 至 1993-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3120302
• 关键词: anemia; anions; ankyrins; antibody; band 3 protein; cellular pathology; chemical binding; congenital blood disorder; cytoskeletal proteins; human population genetics; human subject; immunoglobulin G; ion transport; laboratory rabbit; macrophage; mass spectrometry; molecular pathology; protein sequence; protein structure

The purpose of the proposed study is to investigate hereditary defects in band 3, the major anion transport protein. We have discovered two unique defects in band 3. One of the defects, "slow migrating band3", is characterized by a band 3 molecule that migrates in gels as though it were Mr 98,500 instead of Mr approximately 95,000 as does normal band 3. Thus, it behaves as though there were an addition of Mr approximately 3,500 to the band 3 molecule. The other defect is "fast-migrating band 3" which migrates in gels at Mr=92,000, as though there were a deletion approximately 2,000-3,000 rom the band 3 molecule. Cells from individuals with the latter defect have a significantly shortened lifespan and behave in many ways like old red blood cells. The specific aims of this project are to (1) determine what functional defects are associated with band 3 structural defects; (2) define the structural defects in terms of band 3 domains and amino acid sequence; (3) determine whether the structural changes in band 3 affect its interaction with other membrane and cytoskeletal proteins and whether cytoskeletal proteins are normal; (40 determine the mode of inheritance of band 3 defects in kindreds; (5) characterize antibodies bound to red cells with band 3 anomalies; and (6) identify other defects in band 3 in the population. Band 3 will be characterized by gel electrophoresis before and after treatment with endoglycosidase F to remove carbohydrate groups. The number of IgG molecules per cell will be a determined using a DIDS binding assay. IgG binding studies will be performed. If there are abnormal amounts of IgG on red cells, IgG will be eluted and its specificity determined. Anion transport studies will be performed as a test of band 3 function, and ankyrin binding studies will used to test band 3 interaction with ankyrin. Band 3 proteins that are abnormal as determined by these assays will be analyzed by one and two dimensional peptide mapping and fast atom bombardment mass spectroscopy, and the amino acid sequence of the abnormal segment will be determined. Cytoskeletal proteins will be screened by gel electrophoresis and binding assays to determine whether their interactions with each other are normal. It is anticipated that these studies will provide insights into the structure and function of normal band 3 as well as into the molecular basis of some anemias.

拟议的研究的目的是调查遗传缺陷 带3，主要阴离子转运蛋白。我们发现了两个独特的 乐队3中的缺陷。一个缺陷之一，“慢迁移band3”是 以3个分子为特征，该分子在凝胶中迁移 是MR 98,500而不是MR大约95,000，正常带3。 因此，它的行为好像MR大约增加了 3,500到3个分子。另一个缺陷是“快速移民频段3” MR = 92,000的凝胶迁移，好像有删除 大约2,000-3,000 ROM频段3分子。来自个体的细胞 由于后一个缺陷的寿命大大缩短并表现 在许多方面，例如旧红细胞。 该项目的具体目的是（1）确定什么功能 缺陷与3条结构缺陷有关； （2）定义 结构缺陷在带3个结构域和氨基酸序列方面； （3）确定频段3中的结构变化是否影响其 与其他膜和细胞骨架蛋白的相互作用 细胞骨架蛋白正常； （40确定继承方式 幼儿园中的3个缺陷； （5）表征与红色结合的抗体 带有3个异常的细胞； （6）在频段3中确定其他缺陷 人口。 频段3将以凝胶电泳为特征 用内糖苷酶F处理以去除碳水化合物基团。这 每个细胞的IgG分子的数量将是使用DIS确定的 结合测定。将进行IgG结合研究。如果有 红细胞上IgG的异常量，IgG将洗脱，其IgG量 确定的特异性。阴离子运输研究将作为一个 带3功能的测试和arnkyrin结合研究将用于测试 带3与Ankyrin的相互作用。频带3蛋白异常为 由这些测定法确定的将由一维和二维分析 肽映射和快速原子轰击质谱，以及 将确定异常段的氨基酸序列。 细胞骨架蛋白将通过凝胶电泳和结合筛选 确定他们之间相互作用是否相互作用的测定 普通的。预计这些研究将为您提供见识 正常带3以及分子的结构和功能 一些贫血的基础。

项目成果

Alteration in membrane protein band 3 associated with accelerated erythrocyte aging.

膜蛋白带 3 的改变与加速红细胞老化有关。

• DOI: 10.1073/pnas.86.15.5834
• 发表时间: 1989
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Kay,MM;Flowers,N;Goodman,J;Bosman,G
• 通讯作者: Bosman,G