Phospholipid dynamics and lipolysis in membrane models

膜模型中的磷脂动力学和脂肪分解

• 批准号: 8461140
• 负责人: JOSEPH HAJDU
• 金额: $ 10.49万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8461140
• 关键词: 1,2-diacylglycerol; Arachidonic Acids; Behavior; Biological; Biological Assay; Biological Markers; Biological Process; Biophysics; Cell membrane; Detection; Diglycerides; Disease; Energy Transfer; Enzymatic Biochemistry; Enzymes; Fluorescence; Fluorescence Resonance Energy Transfer; Health; Hydrocarbons; Hypersensitivity; Inflammation; Label; Learning; Lipids; Lipolysis; Lysophosphatidylcholines; Measurement; Mediating; Membrane; Membrane Lipids; Membrane Proteins; Membrane Structure and Function; Metabolic; Methods; Micelles; Monitor; Phospholipase; Phospholipase A2; Phospholipids; Play; Preparation; Property; Proteins; Reaction; Reporter; Research; Role; Series; Signal Transduction; Site; Structure; Tail; Testing; Time; Vesicle; analog; design; fluorophore; in vivo; membrane model; new technology; response; self assembly

DESCRIPTION (provided by applicant): Phospholipids have a key role in contributing to the physicochemical properties of biological membranes. Because of the dynamic character of lipid-lipid and lipid-protein interactions in membranes and membrane mediated biological processes, characterization of the structure and dynamics of phospholipids in self-assembly and in response to addition of metabolites (e.g., other membrane components) is an important step toward understanding membrane biophysics as it relates to cell membrane function and signal transduction. The proposed research aims at characterization of the organization, structure and dynamic behavior of phospholipids in vesicle bilayers and micelles as membrane models using a series of newly designed double- labeled phospholipid analogues with strategically placed (site-directed) fluorescent reporter groups. The spectroscopic labels will be introduced (i) at each of the two fatty acyl chain-ends to characterize chain-chain interactions, and (ii) at the headgroup and each acyl chain, one at a time, to determine the distance / orientation between the polar headgroup and each one of the hydrophobic tails. These probes will be incorporated into vesicles and micelles of defined compositions and physicochemical parameters. Measurements of fluorescence resonance energy transfer (FRET) will be used to characterize the probes. The proposed hypothesis is that determination of the distance and orientation between the constituent fluorescent groups of phospholipid molecules in these synthetic analogues will provide a new technology to determine the structure and dynamics of phospholipid behavior in membranes. In contrast to the commonly used double- labeled fluorescent phospholipid analogues that are almost exclusively used for real-time spectroscopic assay of phospholipases by monitoring the loss of intramolecular fluorescence quenching, the spectroscopic probes here introduced are designed to be used as true "spectroscopic rulers" to determine the distance and orientation of the fluorescent groups in the phospholipid molecule by direct measurement of the FRET efficiency between the labels. Elucidation of phospholipid structure and dynamics in membranes and the way they interact with other membrane lipids and proteins will provide important information toward better understanding membrane behavior in health and disease.

性状（由申请人提供）：磷脂在促进生物膜的理化性质方面具有关键作用。由于膜和膜介导的生物过程中脂质-脂质和脂质-蛋白质相互作用的动态特征，磷脂在自组装中和响应于代谢物（例如，其他膜组分）是理解膜生物物理学的重要一步，因为它涉及细胞膜功能和信号转导。拟议的研究旨在表征的组织，结构和动态行为的磷脂在囊泡双层和胶束作为膜模型，使用一系列新设计的双标记的磷脂类似物与战略性地放置（定点）荧光报告基团。光谱标记将被引入（i）在两个脂肪酰基链末端的每一个处以表征链-链相互作用，和（ii）在头基和每个酰基链处，一次一个，以确定极性头基和每个疏水尾部之间的距离/方向。这些探针将被纳入囊泡和胶束的定义的组成和物理化学参数。荧光共振能量转移（FRET）的测量将用于表征探针。所提出的假设是，确定这些合成类似物中磷脂分子的组成荧光基团之间的距离和方向将提供一种新的技术来确定膜中磷脂行为的结构和动力学。与通常使用的双标记荧光磷脂类似物（其几乎专门用于通过监测分子内荧光淬灭的损失来进行磷脂酶的实时光谱测定）相反，这里介绍的光谱探针被设计成用作真正的“光谱标尺”。通过直接测量标记之间的FRET效率来确定磷脂分子中荧光基团的距离和取向。阐明膜中的磷脂结构和动力学以及它们与其他膜脂质和蛋白质相互作用的方式将为更好地理解健康和疾病中的膜行为提供重要信息。