MOLECULAR STRUCTURE OF BIOLOGICAL MOLECULES

生物分子的分子结构

基本信息

批准号：
6576891
负责人：
DONALD M. ENGELMAN
金额：
$ 17.42万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-01 至 2003-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6576891
关键词：
calorimetry chemical models circular dichroism crystallization lipid bilayer membrane low angle X ray diffraction analysis membrane proteins nuclear magnetic resonance spectroscopy phospholamban polymerase chain reaction protein engineering protein folding protein structure function site directed mutagenesis structural biology

项目摘要

The biological importance of membrane proteins has been underscored by the discovery that at least 25% of all genes in genomic databases have putative transmembrane helices. Yet, there are few structure for membrane proteins, largely because it is hard to create a soluble, stable state for study. We propose to explore alternative methods for structure determination for membrane proteins. We have succeeded in a re-design of phospholamban, a pentameric membrane protein, leading to a stable, pentameric, water-soluble structure that gives long-range NOE signals in NMR. We will use this molecule as a lead compound to explore the principles of stability and aqueous solubility for re-engineering membrane proteins. We will examine the structure of selected signs by NMR and attempt and attempt crystallization. As an alternative to the use of re-design, we will examine the properties of new class of surfactant, polymeric amphiphiles, which has considerable advantages over conventional surfactants, and may provide a new direction for NMR and/or crystallography. We will use solution scattering to measure the thickness of membranes and bilayers of lipids from them. We will continue our use of solution scattering to examine issues of oligomerization and folding of soluble and membrane proteins.

基因组数据库中至少 25% 的基因具有假定的跨膜螺旋，这一发现强调了膜蛋白的生物学重要性。然而，膜蛋白的结构很少，很大程度上是因为很难创造可溶的、稳定的状态用于研究。我们建议探索膜蛋白结构测定的替代方法。我们成功地重新设计了受磷蛋白（一种五聚体膜蛋白），形成了稳定的五聚体水溶性结构，可在 NMR 中提供长程 NOE 信号。我们将使用该分子作为先导化合物来探索重新设计膜蛋白的稳定性和水溶性原理。我们将通过核磁共振检查所选标志的结构并尝试结晶。作为重新设计的替代方案，我们将研究新型表面活性剂——聚合两亲物的性质，它比传统表面活性剂具有相当大的优势，并可能为核磁共振和/或晶体学提供新的方向。我们将使用溶液散射来测量它们的膜和脂质双层的厚度。我们将继续使用溶液散射来检查可溶性和膜蛋白的寡聚和折叠问题。