Effect of Apolipoprotein Structural Adaptability

载脂蛋白结构适应性的影响

• 批准号: 8465891
• 负责人: ROBERT O'Mara RYAN
• 金额: $ 37.82万
• 依托单位: CHILDREN'S HOSPITAL & RES CTR AT OAKLAND
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8465891
• 关键词: Adopted; Affect; Age of Onset; Agonist; Alleles; Alzheimer' s Disease; Amino Acids; Apolipoprotein E; Apolipoproteins; Binding; Biological; Biological Process; Calorimetry; Cardiovascular Diseases; Cell Culture Techniques; Cell Surface Receptors; Cholesterol Homeostasis; Complex; Coronary Arteriosclerosis; Culture Media; Data; Defect; Disease; Disease susceptibility; Experimental Designs; Family; Family member; Generations; Goals; Health; Heteronuclear NMR; Homeostasis; Isotope Labeling; Knowledge; LDL-Receptor Related Proteins; Lead; Ligand Binding; Ligands; Ligation; Lipid Binding; Lipids; Lipoproteins; Low Density Lipoprotein Receptor; Low-Density Lipoproteins; Maintenance; Mediating; Metabolic; Metabolism; Modeling; Molecular; Molecular Conformation; Mutation; N-terminal; NMR Spectroscopy; Nature; Neurodegenerative Disorders; Pathway interactions; Plasma; Predisposition; Protein Isoforms; Proteins; Recombinants; Regulation; Research; Resolution; Risk; Role; Shapes; Site; Structural Models; Structure; Surface Plasmon Resonance; Testing; Therapeutic Agents; Time; Titrations; X-Ray Crystallography; apolipoprotein E-3; apolipoprotein E-4; base; design; disorder prevention; human subject; improved; insight; interest; lipid metabolism; member; novel strategies; particle; public health relevance; receptor; receptor binding; research study; stable isotope; three dimensional structure

DESCRIPTION (provided by applicant): Description The long-term goal of our research is to understand how molecular interactions between lipoproteins and cell surface receptors affect lipid metabolism and disease susceptibility. Knowledge of the metabolic roles of apolipoprotein (apo) E and low-density lipoprotein receptor (LDLR) family members will be combined with available structural information in the design of an experimental strategy to dissect determinants of a productive binding interaction. A novel strategy, termed expressed protein ligation, will be employed to introduce stable isotopes into a specific, predetermined, region of apoE that is essential for LDLR binding. The research to be pursued includes three specific aims. 1) Segmental isotope labeled apoE3-N-temrinal domain, in complex with lipid, will be analyzed by multidimensional heteronuclear NMR spectroscopy in experiments designed to distinguish between alternate structural models of the lipid-bound apoE. 2) The contact sites between apoE and the LDLR will be characterized. It is hypothesized that analysis of segmental isotope labeled apoE3-NT7DMPC in complex with a functional LDLR mini-receptor will yield molecular details of their binding interaction. 3) The interaction of apoE with LDL receptor related protein 6 (LRP6) will be studied. The hypothesis that LDL-A repeats located near the transmembrane spanning sequence of LRP6 are involved in apoE ligand binding will be evaluated. In addition, the postulate that a mutation in LRP6 (R611C), associated with coronary artery disease in human subjects, causes a defect in pH dependent apoE ligand release will be tested. The results of these experiments will extend knowledge of the LDLR family, apoE-mediated lipoprotein metabolism and regulation of plasma lipid homeostasis. Based on the fact that aberrations in the LDLR pathway are positively correlated to onset of cardiovascular disease and apoE manifests isoform-specific susceptibility to neurodegenerative diseases, we anticipate that new knowledge gained from these studies will provide insight into molecular mechanisms that regulate key metabolic processes in health and disease.

描述（由申请人提供）： 描述 我们研究的长期目标是了解脂蛋白和细胞表面受体之间的分子相互作用如何影响脂质代谢和疾病易感性。对载脂蛋白 (apo) E 和低密度脂蛋白受体 (LDLR) 家族成员的代谢作用的了解将与现有的结构信息相结合，设计实验策略，以剖析有效结合相互作用的决定因素。一种称为表达蛋白连接的新策略将用于将稳定同位素引入 apoE 的特定预定区域，该区域对于 LDLR 结合至关重要。要进行的研究包括三个具体目标。 1) 片段同位素标记的 apoE3-N-末端结构域与脂质复合，将在旨在区分脂质结合 apoE 的替代结构模型的实验中通过多维异核 NMR 光谱进行分析。 2) 将表征 apoE 和 LDLR 之间的接触位点。据推测，对与功能性 LDLR 微型受体复合的节段同位素标记的 apoE3-NT7DMPC 的分析将产生它们结合相互作用的分子细节。 3）研究apoE与LDL受体相关蛋白6（LRP6）的相互作用。将评估位于 LRP6 跨膜序列附近的 LDL-A 重复序列参与 apoE 配体结合的假设。此外，还将测试与人类受试者冠状动脉疾病相关的 LRP6 (R611C) 突变导致 pH 依赖性 apoE 配体释放缺陷的假设。这些实验的结果将扩展对 LDLR 家族、apoE 介导的脂蛋白代谢和血浆脂质稳态调节的认识。基于 LDLR 通路的畸变与心血管疾病的发病呈正相关，并且 apoE 表现出对神经退行性疾病的异构体特异性易感性，我们预计从这些研究中获得的新知识将有助于深入了解调节关键代谢过程的分子机制。健康和疾病。