Apolipoprotein A-V: A Functional Proteomics Study

载脂蛋白 A-V：功能蛋白质组学研究

基本信息

批准号：
7905068
负责人：
ROBERT O'Mara RYAN
金额：
$ 40万
依托单位：
CHILDREN'S HOSPITAL & RES CTR AT OAKLAND
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7905068
关键词：
Adenovirus Vector Adenoviruses Adverse effects Affect American Amino Acid Substitution Apolipoproteins Apolipoproteins A Asian Americans Binding Binding Proteins Binding Sites C-terminal Cardiovascular Diseases Cell Culture Techniques Cell Surface Proteins Characteristics Charge Development Diagnosis Disease Dominant-Negative Mutation Elements Endothelial Cells Family member Gene Transfer Genes Genetic Polymorphism Glycosylphosphatidylinositols Goals Grant Heparan Sulfate Proteoglycan High Density Lipoproteins Home environment Homeostasis Hypertriglyceridemia In Vitro Indium Intervention Knockout Mice Knowledge Length Lipids Lipoprotein Binding Lipoproteins Low Density Lipoprotein Receptor Maintenance Measures Mediating Metabolic Metabolic syndrome Metabolism Molecular Mus N-terminal Non-Insulin-Dependent Diabetes Mellitus Obesity Phenotype Physiological Plasma Population Prevention Property Protein Region Proteomics Regulation Relative (related person)Reporting Research Risk Factors Role Structure Testing Transgenic Mice Transgenic Organisms Triglyceride Metabolism Triglycerides Variant apolipoprotein C-II base density design disulfide bond heart disease risk heparin receptor hepatoma cell human population study human subject improved in vitro testing in vivo lipid transport lipoprotein lipase member mouse model mutant prevent public health relevance receptor binding research study

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand the metabolic regulation of plasma triglyceride (TG). Whereas factors such as lipoprotein lipase and apolipoproteins C-II / C-III are known to modulate plasma TG levels, accumulating evidence suggests the recently discovered apolipoprotein A-V (apoA-V) functions in lipid transport and maintenance of plasma TG homeostasis. To elucidate the molecular basis of apoA-V's physiological effects on plasma TG a combination of in vitro and in vivo studies will be performed. Adenovirus mediated gene transfer into apoa5 (-/-) mice will be performed to evaluate in vivo the ability of specific apoA-V variants to influence plasma TG levels and lipoprotein metabolism. Evidence indicates apoA-V binds heparan sulfate proteoglycans, low-density lipoprotein receptor family members and the endothelial cell surface protein glycosylphosphatidylinositol high-density lipoprotein binding protein 1 via a positively charged sequence motif (residues 186-227). Aim 1 will test the hypothesis that this region of the protein is critical for manifestation of the TG modulation properties of apoA-V in vivo. In Aim 2 a variant form of apoA-V correlated with hypertriglcyeridemia (HTG) that results from a c.553G>T SNP will be characterized for intra- molecular disulfide bond formation, heparin / receptor binding interactions and in vivo TG modulation capability. In Aim 3 the hypothesis that the C-terminal domain of apoA-V is necessary and sufficient for TG modulation in vivo will be tested. In addition, in APOA5 transgenic mice, the ability of the NT domain to mimic effects on lipoprotein metabolism seen in human subjects harboring similar truncated forms of apoA-V will be assessed. In Aim 4 the effect of apoA-V lipid droplet association on TG metabolism will be studied in stably transfected hepatoma cells. Knowledge gained will provide a molecular explanation for the relationship between apoA-V and plasma TG levels and will be useful in the design of strategies to diagnose, treat and/or prevent HTG and related disorders. PUBLIC HEALTH RELEVANCE: Statement of Relevance Hypertriglyceridemia is an independent risk factor for cardiovascular disease and is a recognized contributor to development of the metabolic syndrome. Increased understanding of factors that regulate plasma triglyceride levels should provide new opportunities for intervention. Improvements in diagnosis, prevention and/or treatment could have a major impact on the large population of Americans at risk for heart disease, obesity and Type 2 diabetes.

描述（由申请人提供）：我们研究的长期目标是了解血浆甘油三酸酯（TG）的代谢调节。尽管已知脂蛋白脂肪酶和载脂蛋白C-III等因素可以调节血浆TG水平，但积累的证据表明，最近发现的载脂蛋白A-V（APOA-V）在脂质转运和血浆TG稳态的维持中的功能。为了阐明ApoA-V对血浆Tg的生理影响的分子基础，将进行体外和体内研究的组合。将进行腺病毒介导的基因转移到ApOA5（ - / - ）小鼠中，以评估体内特定APOA-V变体影响等离子体TG水平和脂蛋白代谢的能力。有证据表明，ApoA-V结合硫酸肝素蛋白聚糖，低密度脂蛋白受体家族成员和内皮细胞表面蛋白糖基磷脂酰磷脂辛醇高密度脂蛋白结合蛋白1通过呈阳性电荷序列序列基序（残基186-227）。 AIM 1将检验以下假设：蛋白质的该区域对于体内ApoA-V的TG调制特性至关重要。在AIM 2中，APOA-V的变体形式与高三环囊肿（HTG）相关，该变体是由C.553G> t SNP引起的，它将用于分子内二硫化物键的形成，肝素 /受体结合相互作用和VIVO TG调节能力。在AIM 3中，将测试ApoA-V的C末端结构域，足以在体内进行TG调节。此外，在ApoA5转基因小鼠中，将评估NT结构域对在具有类似截短形式的APOA-V的人类受试者中观察到的对脂蛋白代谢的影响的能力。在AIM 4中，将在稳定转染的肝癌细胞中研究ApoA-V脂质液滴关联对TG代谢的影响。获得的知识将为ApoA-V与等离子体TG水平之间的关系提供分子解释，并将在设计诊断，治疗和/或预防HTG和相关疾病的策略方面有用。公共卫生相关性：相关性高甘油三酯血症是心血管疾病的独立危险因素，并且是代谢综合征发展的公认贡献者。对调节血浆甘油三酸酯水平的因素的了解增加应为干预提供新的机会。诊断，预防和/或治疗的改善可能会对患有心脏病，肥胖和2型糖尿病风险的大量美国人产生重大影响。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The carboxyl-terminal segment of apolipoprotein A-V undergoes a lipid-induced conformational change.

DOI：
10.1021/bi1005859
发表时间：
2010-06-15
期刊：
BIOCHEMISTRY
影响因子：
2.9
作者：
Mauldin, Kasuen;Lee, Brian L.;Oleszczuk, Marta;Sykes, Brian D.;Ryan, Robert O.
通讯作者：
Ryan, Robert O.

Apolipoprotein A-V associates with intrahepatic lipid droplets and influences triglyceride accumulation.