STRUCTURAL ANALYSIS OF LISL A WD REPEAT PROTEIN

LISL A WD 重复蛋白的结构分析

基本信息

批准号：
2797160
负责人：
EVA J. NEER
金额：
$ 2.54万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-09-30 至 1999-09-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2797160
关键词：
G protein animal tissue crosslink dimer enzyme activity hydrolase intermolecular interaction microtubules nucleic acid repetitive sequence platelet activating factor protein biosynthesis protein structure function

项目摘要

DESCRIPTION This research proposes to characterize the LIS1 protein with the specific goal of studying the role of the WD repeats in mediating the ability of LIS1 to form protein complexes with itself and other protein subunits. Many proteins containing WD repeats appear to participate in processes involving multi-subunit protein complexes. Previous studies within the US investigator's lab had suggested that the WD repeats in the b subunits of G proteins were important for controlling the reversible formation of G protein trimers and thus mediating the abilities of the different G protein subunits to interact with different effectors and modulating G protein function. The more recent solution of the crystal structure of the Gb subunit shows that the WD repeats participate in the formation of a seven-bladed b propeller structure and are comprised of secondary structural elements consistent with theoretical predictions made based on the analysis of a large library of WD repeat containing proteins. Initial mutagenesis studies within the US lab have tested the contributions of this seven-bladed propeller structure to various aspects of G protein function. This FIRCA application proposes to employ similar approaches to study the function of WD repeats in LIS1 function. The hypothesis being tested is that the presence of WD repeats serve a structural/function relationship common to the widely diverse group of WD-containing proteins. Specifically, the aims are: 1. To define the regions of LIS1 that specify heterodimer or homodimer formation; 2. To define the regions of LIS1 that specify trimer formation with Platelet-activating factor (PAF) acetylhydrolase (AH); 3. To map regions of LIS1 that specify interaction with microtubules by mutational analysis.

描述本研究旨在用特定的特性来表征 LIS1 蛋白研究 WD 重复序列在介导 LIS1 能力中的作用的目标与自身和其他蛋白质亚基形成蛋白质复合物。许多含有 WD 重复序列的蛋白质似乎参与了以下过程：多亚基蛋白质复合物。之前在美国的研究研究人员的实验室表明 WD 在 G 的 b 亚基中重复蛋白质对于控制 G 的可逆形成很重要蛋白质三聚体，从而介导不同 G 蛋白的能力亚基与不同效应子相互作用并调节 G 蛋白功能。 Gb晶体结构的最新解决方案亚基显示 WD 重复参与形成七叶b型螺旋桨结构由二级结构组成与基于分析的理论预测一致的要素包含蛋白质的大型 WD 重复文库。初始诱变美国实验室的研究测试了这种七叶片的贡献螺旋桨结构对 G 蛋白功能的各个方面都有影响。这个FIRCA 申请建议采用类似的方法来研究 WD 在 LIS1 功能中重复。正在检验的假设是 WD 重复序列的存在提供了常见的结构/功能关系包含 WD 的蛋白质种类繁多。具体而言，目标是： 1. 定义指定异源二聚体或同二聚体形成； 2. 定义指定三聚体的 LIS1 区域与血小板激活因子（PAF）乙酰水解酶（AH）形成； 3. 到通过突变指定与微管相互作用的 LIS1 的映射区域分析。