NMR of basic-aromatic clusters

碱性芳香族簇的NMR

• 批准号: 6704789
• 负责人: STEVEN Owen SMITH
• 金额: $ 26.34万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6704789
• 关键词: biological signal transduction; caveolins; cell differentiation; cell membrane; cell proliferation; cholesterol; conformation; fluorescence resonance energy transfer; lipid bilayer membrane; membrane proteins; microscopy; nuclear magnetic resonance spectroscopy; phosphatidylinositols; phosphorylation; protein binding; protein sequence; protein structure function

DESCRIPTION (provided by applicant): The proposed research is to establish the structural basis for how basic-aromatic clusters in membrane-associated proteins are able to sequester specific lipids into lateral membrane domains, how they induce membrane curvature, and how they allow specific peptides to cross cell membranes. The proposal is focused primarily on the MARCKS protein, but also investigates the structure and binding of a wide range of additional basic-aromatic clusters in order to establish how membrane binding and penetration depends on protein sequence. The specific aims of the proposal are to determine 1) the location and conformation of the MARCKS (151-175) effector domain in membrane bilayers, 2) how the protein sequence determines the depth of penetration into bilayers, 3) whether and how basic-aromatic clusters sequester cholesterol, and 4) the role of basic-aromatic clusters in membrane curvature and peptide penetration. The research relies primarily on magic angle spinning NMR measurements of membrane multilayers and solution NMR of membrane bicelles. The NMR approaches are complemented by fluorescence techniques to investigate binding and membrane interactions. Basic-aromatic clusters may be a common mechanism for sequestering phosphoinositides and cholesterol in lateral domains. As a result, their importance in human health is significant. Understanding how the levels of phosphoinositides are regulated is central to signal transduction pathways which are involved in cell growth and differentiation, while understanding how the distribution of cholesterol is controlled in plasma membranes has relevance for cardiovascular and related diseases. The strategy is to progress from simple domains induced by the MARCKS (151-175) peptide to the more complex caveolar domains. The NMR measurements proposed will provide high resolution detail which is not available from crystallographic methods or more traditional solution NMR methods.

描述（由申请人提供）：拟议的研究是为了建立结构性基础，以使膜相关蛋白中的碱性聚集簇能够将特定的特定脂质隔离到侧膜结构域，如何诱导膜曲率以及如何允许特定的肽交叉细胞膜。该提案主要集中在MARCKS蛋白上，但也研究了广泛的其他碱性芳香族簇的结构和结合，以确定膜结合和穿透性如何取决于蛋白质序列。 The specific aims of the proposal are to determine 1) the location and conformation of the MARCKS (151-175) effector domain in membrane bilayers, 2) how the protein sequence determines the depth of penetration into bilayers, 3) whether and how basic-aromatic clusters sequester cholesterol, and 4) the role of basic-aromatic clusters in membrane curvature and peptide penetration.该研究主要依赖于膜多层的魔法旋转NMR测量和膜双层溶液NMR。 NMR方法通过荧光技术补充，以研究结合和膜相互作用。 碱性芳香族簇可能是隔离磷酸肌醇和胆固醇在侧域中的常见机制。结果，它们在人类健康中的重要性非常重要。了解如何调节磷酸肌醇的水平与与细胞生长和分化有关的信号转导途径至关重要，同时了解质膜中胆固醇的分布与心血管及相关疾病的相关性。该策略是从马克克（Marcks）（151-175）肽诱导的简单结构域到更复杂的洞穴结构域的发展。提出的NMR测量结果将提供高分辨率细节，从晶体学方法或更传统的解决方案NMR方法可用。