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酯（TG）的代谢调节。尽管已知脂蛋白脂肪酶和载脂蛋白 C-II / C-III 等因子可调节血浆 TG 水平，但越来越多的证据表明，最近发现的载脂蛋白 A-V (apoA-V) 在脂质转运和维持血浆 TG 稳态中发挥作用。为了阐明apoA-V对血浆TG的生理作用的分子基础，将进行体外和体内研究的结合。将通过腺病毒介导的基因转移至 apoa5 (-/-) 小鼠中，以评估体内特定 apoA-V 变体影响血浆 TG 水平和脂蛋白代谢的能力。有证据表明 apoA-V 通过带正电荷的序列基序（残基 186-227）结合硫酸乙酰肝素蛋白聚糖、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肌醇高密度脂蛋白结合蛋白 1。目标 1 将检验以下假设：该蛋白质区域对于 apoA-V 体内 TG 调节特性的表现至关重要。在目标 2 中，与由 c.553G>T SNP 产生的高甘油三酯血症 (HTG) 相关的 apoA-V 变体形式将被表征为分子内二硫键形成、肝素/受体结合相互作用和体内 TG 调节能力。在目标 3 中，将测试 apoA-V 的 C 末端结构域对于体内 TG 调节是必要且充分的假设。此外，在 APOA5 转基因小鼠中，将评估 NT 结构域模拟对脂蛋白代谢的影响的能力，这种影响在具有类似截短形式的 apoA-V 的人类受试者中观察到。在目标 4 中，将在稳定转染的肝癌细胞中研究 apoA-V 脂滴结合对 TG 代谢的影响。获得的知识将为apoA-V和血浆TG水平之间的关系提供分子解释，并将有助于设计诊断、治疗和/或预防HTG和相关疾病的策略。公共卫生相关性：相关性声明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生的公认因素。对调节血浆甘油三酯水平的因素的更多了解应该为干预提供新的机会。诊断、预防和/或治疗方面的改进可能会对大量面临心脏病、肥胖症和 2 型糖尿病风险的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.