Project 3

项目3

• 批准号: 8151962
• 负责人: CHARLES R SANDERS
• 金额: $ 46.91万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8151962
• 关键词: Biological; Cholesterol; Detergents; Disease; Ethers; Head; Integral Membrane Protein; Link; Lipids; Lysophospholipids; Measurement; Membrane; Membrane Proteins; Methods; Micelles; Mole the mammal; Molecular Chaperones; Molecular Conformation; Organism; Phosphotransferases; Polymers; Preparation; Relative (related person); Relaxation; Resources; Sampling; Solubility; Solutions; Source; Staging; Streptococcus mutans; Structure; System; Testing; Work; base; cholesterol analog; design; dimer; human PMP22 protein; membrane model; mimetics; monomer; novel; protein structure; restraint; structural biology; structural genomics; surfactant

3.3. Background and Significance 3.3.1. There is a compelling need for novel model membrane media that are optimized for MP structural biology. The vast majority of MP structures determined to date by either NMR or crystallographic methods were determined using classical detergent as the model membrane media. Unfortunately, even mild detergents destabilize MPs relative to their stability in native membranes. Moreover, many of the mildest detergents (such as the alkyl maltoside) routinely fail to yield high quality NMR spectra. More native-like media, such as detergent-lipid mixed micelles or bicelles sometimes offer advantages, but also have drawbacks. Moreover, while membranes from higher organisms almost always contain a considerable mole fraction of cholesterol, very little work has been carried out to establish NMR-compatible media that contain cholesterol, in part because of its very low solubility in detergent solutions. It is clear that there is a need to expand the range of membrane-mimetic media that are available. We propose to design, synthesize (through Core B) and test new surfactants for use in structural biological studies of MPs. We also will test surfactants and cholesterol analogs that have recently been commercialized but not yet well tested.

3.3.背景及意义 3.3.1.迫切需要针对 MP 结构生物学进行优化的新型模型膜介质。迄今为止，通过 NMR 或晶体学方法确定的绝大多数 MP 结构都是使用经典洗涤剂作为模型膜介质来确定的。不幸的是，即使是温和的去污剂也会使 MP 相对于其在天然膜中的稳定性变得不稳定。此外，许多最温和的洗涤剂（例如烷基麦芽糖苷）通常无法产生高质量的核磁共振谱。更接近天然的介质，例如去污剂-脂质混合胶束或双胶束有时具有优点，但也有缺点。此外，虽然来自高等生物体的膜几乎总是含有相当大的胆固醇摩尔分数，但在建立含有胆固醇的 NMR 兼容介质方面所做的工作却很少，部分原因是其在洗涤剂溶液中的溶解度非常低。显然，需要扩大可用的膜模拟介质的范围。我们建议设计、合成（通过核心 B）和测试新的表面活性剂，用于 MP 的结构生物学研究。我们还将测试表面活性剂 以及最近已商业化但尚未经过充分测试的胆固醇类似物。