Structural Basis of the Anti-Atherogenic Properties of ApoA-I

ApoA-I 抗动脉粥样硬化特性的结构基础

基本信息

批准号：
7596522
负责人：
MICHAEL CANAVAN PHILLIPS
金额：
$ 34.2万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-01-01 至 2013-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7596522
关键词：
ATP-Binding Cassette Transporters Address Adopted Affect Affinity Amino Acids Animals Antiatherogenic Apolipoprotein A-I Apolipoproteins Apolipoproteins A Atherosclerosis Binding Biochemical Biogenesis Biological Assay C-terminal Cells Characteristics Cholesterol Collaborations Coronary Arteriosclerosis Dependovirus Engineering Fibroblasts Goals Hepatocyte Heterogeneity High Density Lipoprotein Cholesterol High Density Lipoproteins Human Human Engineering Hybrids Hydrophobicity Incidence Incubated Lipid Binding Lipids Lipoproteins Measurement Measures Mediating Metabolism Methods Modeling Molecular Molecular and Cellular Biology Mus Mutation N-terminal Particle Size Performance Phospholipids Physiological Plasma Plasma Proteins Play Population Property Protein Engineering Proteins Relative (related person)Research Personnel Risk Role SR-BI receptor Solutions Structure Structure-Activity Relationship Surface Testing Variant base cell type cholesterol transporters design lipid transport macrophage mutant particle physical property premature programs protective effect protein function reverse cholesterol transport synthetic peptide

项目摘要

pprwinpn PROJECT 2: STRUCTURAL BASIS OF THE ANTI-ATHEROGENIC PROPERTIES OF APO A-l The goal of this project is to elucidate the molecular mechanisms underlying the functions of apolipoprotein (apo) A-l in reverse cholesterol transport (RCT). ApoA-l is the major protein of plasma high- density lipoprotein (HDL)and the functions of this molecule underlie the anti-atherogenic properties of the lipoprotein. Specific Aim 1 is to define the key properties of the two tertiary structure domains of the apoA-l molecule, and to better define lipid-free human apoA-l structure in dilute solution at physiological concentrations. The influence of the properties of the two apoA-l structural domains on the affinity and mechanism of lipid binding will also be investigated. These objectives will be accomplished by using engineered apoA-l molecules and a range of physical-biochemical methods. Specific Aim 2 is to establish the mechanistic basis for the biogenesis of heterogeneous nascent HDL particles via the ATP-binding cassette transporter Al (ABCA1)-mediated efflux of cellular lipids to apoA-l. The role of apoA-l structure and influence of cell type will be determined by measuring the efflux of phospholipid and cholesterol molecules from macrophages, fibroblasts and liver cells growing in culture to engineered apoA-l molecules. Specific Aim 3 is to define the effects of the properties of the tertiary structural domains of apoA-l on the protein's functionality in cholesterol transport using adeno-associated virus-induced expression of natural human apoA-l variants in mice to assess the effects on RCT and atherosclerosis (in collaboration with Project 3). The functionalities of the mouse HDL particles containing the apoA-l mutations in mediating cholesterol transport into and out of cells will be determined in collaboration with Project 1. The incidence of premature coronary artery disease is reduced in human populations with elevated levels of plasma HDL cholesterol and apoA-l. The reasons for this protective effect are not understood fully and this project seeks to uncover the molecular mechanisms underlying the beneficial properties of apoA-l.

pprwinpn 项目 2：APO A-1 抗动脉粥样硬化特性的结构基础该项目的目标是阐明其功能背后的分子机制反向胆固醇转运 (RCT) 中的载脂蛋白 (apo) A-l。 ApoA-l 是血浆高浓度的主要蛋白质密度脂蛋白（HDL）和该分子的功能是其抗动脉粥样硬化特性的基础脂蛋白。具体目标 1 是定义 apoA-1 的两个三级结构域的关键特性分子，并更好地定义生理学稀溶液中的无脂人 apoA-l 结构浓度。两个apoA-l结构域的性质对亲和力和亲和力的影响脂质结合机制也将得到研究。这些目标将通过使用来实现工程apoA-l分子和一系列物理生化方法。具体目标 2 是建立通过 ATP 结合产生异质新生 HDL 颗粒的机制基础盒式转运蛋白 A1 (ABCA1) 介导细胞脂质流出至 apoA-l。 apoA-l 结构和作用通过测量磷脂和胆固醇分子的流出来确定细胞类型的影响从培养中生长的巨噬细胞、成纤维细胞和肝细胞到工程化的 apoA-l 分子。具体的目标 3 是确定 apoA-1 三级结构域的特性对蛋白质的影响使用腺相关病毒诱导的自然人表达在胆固醇运输中的功能小鼠中的 apoA-1 变体评估对 RCT 和动脉粥样硬化的影响（与项目 3 合作）。含有apoA-1突变的小鼠HDL颗粒在介导胆固醇方面的功能进出细胞的运输将与项目 1 合作确定。在血压升高的人群中，过早冠状动脉疾病的发生率降低血浆 HDL 胆固醇和 apoA-l 水平。这种保护作用的原因尚不完全清楚该项目旨在揭示 apoA-l 有益特性背后的分子机制。