Improving membrane protein crystallization

改善膜蛋白结晶

基本信息

批准号：
7493752
负责人：
JAMES U BOWIE
金额：
$ 28.26万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-15 至 2010-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7493752
关键词：
Adopted Binding Chimeric Proteins Compatible Crystallization Crystallography Cytoplasmic Tail Data Detergents Diacylglycerol Kinase Dihydrofolate Reductase Disease Engineering Genetic Variation Glucose Transporter Glycophorin A Goals Hand Health Helix (Snails)Judgment Lateral Learning Left Ligand Binding Ligands Medical Membrane Proteins Methods Modification Molecular Conformation Muramidase Needles Numbers Personal Satisfaction Polyhomeotic Polymers Principal Investigator Process Protein Engineering Proteins Range Rate Refractory Research Personnel Resolution SAM Domain Screening procedure Shapes Sodium Structure Surface Tertiary Protein Structure Testing Thick Variant Work base ear helix high throughput screening improved insight new technology novel novel strategies polymerization programs protein folding protein function protein structure research study rhomboid scaffold

项目摘要

DESCRIPTION (provided by applicant): One the primary barriers to membrane protein structure determination by x-ray crystallography is obtaining high quality crystals. We propose two new technologies to improve our ability to obtain crystals of membrane proteins. Method I: We propose a general, high-throughput method to screen for compounds that bind to a target membrane protein. The compounds will stabilize and possibly rigidify the protein, improving the chances of crystallization. The screening method involves a simple test for protein stability. Method II: We proposed to utilize a polymerizing protein module to drive the crystallization of an attached membrane protein. We have demonstrated that the protein module can induce the crystallization of a wide variety of soluble proteins, including those otherwise refractory to crystallization. Moreover, the module is compatible with detergents and preliminary tests indicate that it can drive the crystallization of a transmembrane helix. We plan to further optimize this protein module and test its ability to produce crystals of a range of membrane proteins. Health Relevance: Many diseases are caused by aberrant protein function. Protein function is intimately tied to the elaborate three-dimensional shapes that proteins adopt. It is therefore an important medical goal to learn protein structures so that we can understand how they work and how we can intervene when their functions go awry in disease. This proposal seeks to increase the rate at which we can obtain this critical structural information.

描述（由申请人提供）：通过X射线晶体学测定膜蛋白结构的主要障碍是获得高质量的晶体。我们提出了两种新技术，以提高我们获得膜蛋白晶体的能力。方法I：我们提出了一种一般的高通量方法来筛选与靶膜蛋白结合的化合物。这些化合物将稳定并可能稳定蛋白质，从而提高结晶的机会。筛选方法涉及对蛋白质稳定性的简单测试。方法II：我们提议利用聚合蛋白模块来驱动附着的膜蛋白的结晶。我们已经证明，蛋白质模块可以诱导多种可溶性蛋白的结晶，包括那些对结晶的难治性的蛋白质。此外，该模块与洗涤剂兼容，初步测试表明它可以驱动跨膜螺旋的结晶。我们计划进一步优化该蛋白质模块，并测试其产生各种膜蛋白晶体的能力。健康相关性：许多疾病是由异常蛋白功能引起的。蛋白质功能与蛋白质所采用的精细的三维形状密切相关。因此，学习蛋白质结构是一个重要的医学目标，以便我们可以理解它们的工作方式以及当它们的功能在疾病中脱颖而出时如何干预。该提案旨在提高我们获得此关键结构信息的速度。