Membrane Control of Protein-Protein Contact. Simulation Based on Phase Diagrams

蛋白质-蛋白质接触的膜控制。

• 批准号: 7880973
• 负责人: GERALD William FEIGENSON
• 金额: $ 7.13万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-13 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7880973
• 关键词: 1,2-diacylglycerol; Algorithms; Animals; Behavior; Binding; Buffers; Cell Line; Cell membrane; Cell surface; Cells; Ceramides; Chemicals; Chickens; Cholesterol; Computer Simulation; Coupling; Data; Dependence; Development; Diglycerides; Disease; Drug Combinations; Drug Controls; Ensure; Fibroblasts; Fluorescence Microscopy; Fluorescence Resonance Energy Transfer; Goals; Gramicidin; Grouping; HIV; HIV-1; Human; Invaded; Laboratories; Lead; Life; Life Cycle Stages; Lipids; Liposomes; Measurement; Measures; Membrane; Microscopic; Modeling; Peptides; Perylene; Phase; Phase II Clinical Trials; Proteins; Research; Research Personnel; Rous sarcoma virus; Signaling Molecule; Temperature; Viral; Viral Matrix Proteins; Viral Proteins; Virus; base; behavior change; programs; protein protein interaction; research study; simulation

DESCRIPTION (provided by applicant): Viruses invade all forms of life, causing diseases in humans including HIV-AIDS. Although the variety of viruses is daunting, all enveloped viruses, including HIV-1 and Rous Sarcoma Virus (RSV), must associate with the inner leaflet of plasma membranes. Our laboratories have undertaken a large-scale effort to characterize interactions between membrane components. The goal is to discover which membrane factors exert control over the interactions between membrane-associated viral matrix proteins. The overall approach is to combine experimental measurements, mainly FRET and fluorescence microscopy, with Monte Carlo computer simulations in order to discover all of the significant interaction energies between lipid-lipid, lipid-protein, and protein-protein. Membrane phase behavior and protein-protein interactions are first measured in model liposomes and then in live chicken fibroblast cells. The first specific aim is to develop the experiments and Monte Carlo simulations to describe gramicidin-gramicidin interactions in a simple 3-lipid component mixture containing cholesterol. The second aim is to handle the greatly increased complexity of a fifth component, ceramide or diacylglycerol. This aim includes proposed development of a "lipid buffer" in order to control the chemical potential of cholesterol, ceramide, or diacylglycerol in model or real biomembranes. The third aim is to characterize HIV-1 and RSV matrix protein-protein interactions in a lipid mixture that models the inner leaflet of an animal cell plasma membrane, as a basis for understanding behavior in live cells. The fourth and final aim is to examine HIV-1 and RSV matrix protein-protein interactions at the inner leaflet of the plasma membrane using FRET in live cells, attempting to find the factors that control the interactions, and whether coupling to the outer leaflet is observed. Useful treatments for viral afflictions, in particular HIV-AIDS, are not limited to "all or none" cures. Combinations of drugs, each of which slows down a step of the viral life cycle, have proven useful. The proposed research will lead to detailed understanding of how the membrane components cholesterol, ceramide, and diacylglycerol, all of which can be partially regulated by drugs, control one critical step in the life of the virus - association/assembly at the plasma membrane.

描述（由申请人提供）：病毒侵入所有生命形式，导致人类疾病，包括艾滋病毒/艾滋病。 尽管病毒种类繁多，但所有有包膜病毒，包括 HIV-1 和劳斯肉瘤病毒 (RSV)，都必须与质膜的内层结合。 我们的实验室进行了大规模的努力来表征膜组件之间的相互作用。 目标是发现哪些膜因子对膜相关病毒基质蛋白之间的相互作用进行控制。 总体方法是将实验测量（主要是 FRET 和荧光显微镜）与蒙特卡罗计算机模拟相结合，以发现脂质-脂质、脂质-蛋白质和蛋白质-蛋白质之间的所有重要相互作用能量。 首先在模型脂质体中测量膜相行为和蛋白质-蛋白质相互作用，然后在活鸡成纤维细胞中测量。 第一个具体目标是开发实验和蒙特卡罗模拟来描述短杆菌肽-短杆菌肽在含有胆固醇的简单 3-脂质成分混合物中的相互作用。 第二个目标是处理第五种成分神经酰胺或二酰甘油的复杂性大大增加。 该目标包括提议开发“脂质缓冲剂”，以控制模型或真实生物膜中胆固醇、神经酰胺或二酰甘油的化学势。 第三个目标是表征脂质混合物中的 HIV-1 和 RSV 基质蛋白-蛋白质相互作用，模拟动物细胞质膜的内叶，作为了解活细胞行为的基础。 第四个也是最后一个目标是在活细胞中使用 FRET 检查质膜内层的 HIV-1 和 RSV 基质蛋白-蛋白质相互作用，试图找到控制相互作用的因素，以及是否观察到与外层的耦合。对病毒性疾病、特别是艾滋病毒-艾滋病的有效治疗并不限于“全部或全部”治愈。 药物组合已被证明是有用的，每种药物组合都可以减缓病毒生命周期的一个步骤。 拟议的研究将详细了解膜成分胆固醇、神经酰胺和二酰甘油（所有这些成分都可以部分地受到药物调节）如何控制病毒生命中的一个关键步骤——在质膜上的结合/组装。