Structures, Dynamics, and Functions of Membrane Proteins

膜蛋白的结构、动力学和功能

• 批准号: 10206183
• 负责人: STANLEY J OPELLA
• 金额: $ 51.28万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10206183
• 关键词: Bacterial Proteins; Biochemical; Biology; Carrier Proteins; Crystallization; Detergents; Disease; Drug Metabolic Detoxication; Drug Targeting; Electron Microscopy; Environment; G-Protein-Coupled Receptors; Goals; HIV-1; Hepatitis C virus; Human Genome; Hydrophobicity; Immobilization; Medicine; Membrane; Membrane Proteins; Mercury; Methods; Molecular; Molecular Biology; Molecular Conformation; Mutation; Phospholipids; Physiological; Protein Dynamics; Proteins; Research; Sampling; Structure; System; Technology; Temperature; X-Ray Crystallography; base; chemokine receptor; cost; cryogenics; hydrophilicity; innovation; method development; protein protein interaction

7. Project Summary Membrane proteins are important targets for structure determination. One quarter of the proteins in the human genome are membrane proteins, and the majority of drug targets are membrane proteins. However, both structure determination and dynamics characterization of membrane proteins have lagged because of technological difficulties resulting largely from the phospholipid environment in which the proteins reside; it is highly asymmetric with hydrophobic and hydrophilic components, and it immobilizes the proteins. Consequently, our overarching goal is to develop new and more powerful methods for determining the structures of membrane proteins. This is an essential first step in revealing the bases of their functions. Although considerable progress has been made using solution NMR, X-ray crystallography, and electron microscopy, this has been at the potential cost of distorting the structures through the use of detergent environments and cryogenic temperatures for the protein samples. None of the proteins that we are studying have been crystallized. Thus, the technology gap that we seek to fill is well defined. Innovative features of the research plan include its scientific breadth, which ranges from molecular biology to structure calculations. Several smaller membrane proteins will be used for methods development, including Vpu from HIV-1, p7 from HCV, and mercury transport proteins from the bacterial mercury detoxifications system. They serve the dual purposes of posing interesting biochemical functional questions and serving as tractable systems for developing methods of determining the structures and describing the dynamics of larger membrane proteins, such as our principal target of G-protein coupled receptors (GPCRs). At the conclusion of the research plan, we expect to be able to describe the functions of chemokine receptors. These studies will involve the structures and dynamics of the monomeric proteins, protein-protein interactions, and conformational changes in the proteins. NMR is unique in its ability to characterize global and local dynamics of proteins. Thus, the findings will be highly complementary to parallel studied by x-ray crystallography and electron microscopy. Moreover, NMR is capable of describing structure, dynamics, and interactions of the proteins in their phospholipid bilayer environment under physiological conditions.

7。项目摘要 膜蛋白是结构确定的重要目标。人类中四分之一的蛋白质 基因组是膜蛋白，大多数药物靶标是膜蛋白。但是，这两个 膜蛋白的结构确定和动态表征滞后于 蛋白质所居住的磷脂环境很大程度上造成的技术困难。这是 高度不对称，具有疏水和亲水性成分，并固定蛋白质。 因此，我们的总体目标是开发新的，更强大的方法来确定 膜蛋白的结构。这是揭示其功能基础的重要第一步。 尽管使用溶液NMR，X射线晶体学和电子取得了长足的进步 显微镜，这是通过使用洗涤剂来扭曲结构的潜在成本 蛋白质样品的环境和低温温度。我们没有研究的蛋白质 已经结晶了。因此，我们寻求填补的技术差距是很好的定义。 研究计划的创新特征包括其科学广度，从分子生物学到 结构计算。几种较小的膜蛋白将用于方法开发，包括 来自HIV-1的VPU，HCV的P7和细菌汞解毒的汞转运蛋白 系统。他们达到了提出有趣的生化功能问题的双重目的 用于开发确定结构和描述较大动态的方法的可访问系统 膜蛋白，例如我们的G蛋白耦合受体（GPCR）的主要靶标。 在研究计划的结尾，我们希望能够描述趋化因子受体的功能。 这些研究将涉及单体蛋白，蛋白质蛋白相互作用的结构和动力学， 和蛋白质的构象变化。 NMR在表征全球和本地的能力方面是独一无二的 蛋白质的动力学。因此，发现将与X射线平行研究高度互补 晶体学和电子显微镜。此外，NMR能够描述结构，动力学和 在生理条件下，蛋白质在其磷脂双层环境中的相互作用。