Perilipins and cellular triacyglycerol metabolism

Perilipins 和细胞三酰甘油代谢

• 批准号: 7214083
• 负责人: DAWN L BRASAEMLE
• 金额: $ 27.75万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-15 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214083
• 关键词: 1,2-diacylglycerol; Abbreviations; Address; Adipocytes; Adipose tissue; Affect; Alanine; Amino Acid Sequence; Amino Acids; Aspartic Acid; Binding; Biological Assay; Cardiovascular Diseases; Cells; Chimera organism; Chimeric Proteins; Chinese Hamster; Cholesterol Esters; Condition; Cultured Cells; Cyclic AMP-Dependent Protein Kinases; Diglycerides; Disease; Docking; Drops; Energy Metabolism; Energy-Generating Resources; Enzymes; Fatty Acids; Fatty acid glycerol esters; Fibroblasts; Forskolin; Glutamic Acid; Glutathione S-Transferase; Goals; Green Fluorescent Proteins; Health; Hormonal; Human; Hydrolase; Immunoblotting; Immunofluorescence Microscopy; In Vitro; Incubated; Insecta; Isoproterenol; Lipase; Lipids; Lipolysis; Localized; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Metabolic; Metabolism; Molecular; Monoglycerides; Mutate; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Nucleotides; Obesity; Organelles; Ovary; Patients; Phospholipid Metabolism; Phosphorylation; Play; Point Mutation; Prevalence; Proteins; Proteomics; RNA Interference; Rate; Recombinant Proteins; Recombinants; Regulation; Research; Research Personnel; Role; Serine; Site; Site-Directed Mutagenesis; Surface; Testing; Therapeutic; Tissues; Transferase; Triglycerides; adipophilin; caveolin 1; fatty acid binding protein; fatty acid-binding proteins; feeding; insulin sensitivity; member; perilipin; perilipin A; programs; protein protein interaction; research study; scaffold; sterol esterase; umbelliferyl phosphate

DESCRIPTION (provided by applicant): Obesity is a serious health problem throughout the world, and contributes to an increased prevalence of Type II diabetes, cardiovascular disease, and cancer. Despite a need for therapeutic options to treat obesity, little is known about the mechanisms that regulate fat storage and release from adipocytes. Triacylglycerols (TAG), the body's major storage form of energy, are packaged in lipid droplets in adipocytes that are covered with perilipin A, a protein that plays a critical role in controlling TAG storage and lipolysis to release fatty acids that serve as a major source of energy. The proposed studies will elucidate the molecular mechanisms by which perilipin A coordinates adipocyte TAG metabolism. Preliminary studies have provided evidence that hormone-sensitive lipase (HSL), and CGI-58, a protein important for TAG metabolism, associate with lipid droplets via a mechanism that is responsive to the phosphorylation state of perilipin A. The proposed studies will test the hypotheses that 1) the phosphorylation of perilipin A controls lipolysis through multiple complementary mechanisms, and 2) that perilipin A serves as an organizing center that coordinates the association of TAG catabolic enzymes with lipid droplets in a manner that is responsive to the lipolytic status of the adipocyte. Specifically, we will 1) elucidate the mechanisms by which perilipin A facilitates the docking of HSL on lipid droplets, 2) identify additional mechanisms by which phosphorylated perilipin A coordinates lipolysis that are distinct from its role in facilitating HSL docking, and 3) elucidate the role that perilipin A plays in docking CGI-58 on lipid droplets and the consequences of this functional interaction to TAG metabolism. Intact and mutated perilipins and HSL or CGI-58 will be expressed in cultured cells that lack these proteins followed by assays for TAG storage and lipolysis; additionally, RNAi approaches and in vitro studies of recombinant proteins are proposed. These studies will contribute to a long-term goal of defining the factors on lipid droplets that control adipocyte TAG metabolism.

描述（由申请人提供）：肥胖是全世界一个严重的健康问题，导致 II 型糖尿病、心血管疾病和癌症的患病率增加。尽管需要治疗肥胖的治疗方案，但人们对调节脂肪细胞储存和释放脂肪的机制知之甚少。三酰甘油 (TAG) 是人体能量的主要储存形式，被包装在脂肪细胞中的脂滴中，这些脂肪细胞被周脂蛋白 A 覆盖，周脂蛋白 A 是一种蛋白质，在控制 TAG 储存和脂解以释放作为主要能量来源的脂肪酸方面发挥着关键作用。拟议的研究将阐明 perilipin A 协调脂肪细胞 TAG 代谢的分子机制。初步研究提供的证据表明，激素敏感性脂肪酶 (HSL) 和 CGI​​-58（一种对 TAG 代谢很重要的蛋白质）通过一种对 perilipin A 的磷酸化状态做出反应的机制与脂滴相关联。拟议的研究将检验以下假设：1) perilipin A 的磷酸化通过多种互补机制控制脂肪分解，2) perilipin A 充当组织中心，以响应脂肪细胞脂解状态的方式协调 TAG 分解代谢酶与脂滴的关联。具体来说，我们将1)阐明perilipin A促进HSL在脂滴上对接的机制，2)确定磷酸化perilipin A协调脂肪分解的其他机制，这些机制不同于其在促进HSL对接中的作用，以及3)阐明perilipin A在将CGI-58对接在脂滴上所起的作用 以及这种功能性相互作用对 TAG 代谢的影响。完整和突变的周脂质和 HSL 或 CGI-58 将在缺乏这些蛋白质的培养细胞中表达，然后进行 TAG 储存和脂肪分解测定；此外，还提出了 RNAi 方法和重组蛋白的体外研究。这些研究将有助于确定控制脂肪细胞 TAG 代谢的脂滴因素的长期目标。