FUNCTIONS OF ANKYRINS IN MICE

锚蛋白在小鼠中的功能

• 批准号: 6241851
• 负责人: SAMUEL E LUX
• 金额: $ 30.43万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 1998-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6241851
• 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).

Ankyrins很大，分布广泛，三方蛋白 整体膜转运蛋白，粘合蛋白和受体 基于光谱的膜骨架。 三个伸蛋白被克隆了， 到目前为止的特征是：ANK1（促红细胞Ankyrin），Ank2（脑ankyrin）， 和Ank3（上皮伸蛋白）。 每个伸道蛋白包含一个N末端 “重复”结构域连接到积分膜蛋白，A 中央光谱结合结构域和一个C末端的“调节”域， 其功能没有表征。 ANK3也存在于较小的 缺乏重复域的同工型（ANK3-5KB）。 在巨噬细胞中，ANK3-5KB 仅与晚期溶酶体和溶酶体有关 囊泡。 目前的研究旨在测试氨基链蛋白功能的各个方面 体内，尤其是ANK1是否可以被其他脚踝替换 以及用ANK1代替本地ANK1的后果是什么 调节域的结构变化。 这些问题 将使用靶向基因替代的技术来追求小鼠 胚胎（ES）细胞。 诱导突变对 生存，器官功能和病理，红细胞寿命和 形态学，红细胞膜和膜骨骼组成以及 组织，膜稳定性以及带的功能和机动性 3将进行调查。 其他实验将针对 识别靶向ANK3-5 Kb的序列 识别和克隆靶配体。