FUNCTIONS OF ANKYRINS IN MICE

锚蛋白在小鼠中的功能

• 批准号: 6202237
• 负责人: SAMUEL E LUX
• 金额: $ 30.53万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6202237
• 关键词: ankyrins; band 3 protein; cell growth regulation; embryonic stem cell; erythrocyte membrane; erythropoiesis; gene targeting; laboratory mouse; membrane activity; molecular cloning; protein isoforms; protein structure function; receptor binding; vesicle /vacuole

Ankyrins are large, widely distributed, tripartite proteins that link integral membrane transporters, adhesive proteins and receptors to the spectrin-based membrane skeleton. Three ankyrins have been cloned and characterized so far: Ank1 (erythrocyte ankyrin), Ank2 (brain ankyrin), and Ank3 (epithelial ankyrin). Each ankyrin contains an N-terminal "repeat" domain that attaches ankyrin to integral membrane proteins, a central spectrin-binding domain, and a C-terminal, "regulatory" domain, whose function is not characterized. Ank3 also exists in a smaller isoform (Ank3-5kb)that lacks the repeat domain. In macrophages Ank3-5kb is associated exclusively with late lysosomes and post lysosomal vesicles. The present studies aim to test aspects of ankyrin function in vivo, particularly whether Ank1 can be replaced by the other ankyrins and what the consequences are of replacing native Ank1 with an Ank1 bearing structural changes in the regulatory domain. These questions will be pursued using the technique of targeted gene replacement in mouse embryonal stem (ES) cells. The effects of the induced mutations on survival, organ function and pathology, red cell life span and morphology, red cell membrane and membrane skeletal composition and organization, membrane stability, and the function and mobility of band 3 will be investigated. Additional experiments will be directed at identifying sequences that target Ank3-5 kb to intracellular vesicles and identifying and cloning the target ligand(s).

锚蛋白是大的、分布广泛的三联蛋白，将 整合膜转运蛋白、粘附蛋白和受体 基于血影蛋白的膜骨架。 三个锚蛋白已被克隆 迄今为止的特征：Ank1（红细胞锚蛋白）、Ank2（脑锚蛋白）、 和Ank3（上皮锚蛋白）。 每个锚蛋白含有一个 N 末端 将锚蛋白附着到整合膜蛋白上的“重复”结构域， 中央血影蛋白结合结构域和 C 端“调节”结构域， 其功能未表征。 Ank3 也存在于更小的 缺少重复结构域的异构体 (Ank3-5kb)。 在巨噬细胞中 Ank3-5kb 仅与晚期溶酶体和后溶酶体相关 囊泡。 目前的研究旨在测试锚蛋白功能的各个方面 体内，特别是 Ank1 是否可以被其他锚蛋白取代 用 Ank1 替换原生 Ank1 会产生什么后果 监管领域的结构性变化。 这些问题 将使用小鼠靶向基因替换技术进行研究 胚胎干（ES）细胞。 诱导突变的影响 生存、器官功能和病理、红细胞寿命和 形态、红细胞膜和膜骨架组成以及 组织、膜稳定性以及带的功能和移动性 3将被调查。 额外的实验将针对 鉴定将 Ank3-5 kb 靶向细胞内囊泡的序列，以及 识别和克隆目标配体。