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

• 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
• 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

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：载脂蛋白A-I抗动脉粥样硬化特性的结构基础 该项目的目标是阐明的功能的分子机制， 载脂蛋白（apo）A-I在胆固醇逆向转运（RCT）中的作用。ApoA-I是血浆高脂血症的主要蛋白质， 高密度脂蛋白（HDL）和这种分子的功能是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性质， 分子，并更好地定义在生理条件下稀释溶液中的无脂质人apoA-I结构。 浓度的两个apoA-1结构域的性质对亲和力和亲和力的影响 还将研究脂质结合的机制。这些目标将通过使用 工程化apoA-I分子和一系列物理-生物化学方法。具体目标2是建立 通过ATP结合的异质新生HDL颗粒的生物发生的机制基础 盒式转运蛋白A1（ABCA 1）介导的细胞脂质向apoA-1的流出。apoA-I结构的作用和 细胞类型的影响将通过测量磷脂和胆固醇分子的流出来确定 从培养物中生长的巨噬细胞、成纤维细胞和肝细胞到工程化的apoA-1分子。具体 目的3是确定apoA-1三级结构域的性质对蛋白质的功能的影响， 使用腺相关病毒诱导表达的天然人胆固醇转运功能 apoA-I变体在小鼠中的作用，以评估对RCT和动脉粥样硬化的影响（与项目3合作）。 载脂蛋白A-I突变小鼠高密度脂蛋白颗粒介导胆固醇的功能 将与项目1合作确定进出细胞的运输。 在高血压人群中，早发冠状动脉疾病的发病率降低， 血浆HDL胆固醇和apoA-1水平。这种保护作用的原因还不完全清楚 该项目旨在揭示apoA-1有益特性的分子机制。