Mechanism and physiology of DUF1222-assisted lipase maturation

DUF1222 辅助脂肪酶成熟的机制和生理学

• 批准号: 8269607
• 负责人: Saskia Neher
• 金额: $ 24.5万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8269607
• 关键词: Acids; Adipocytes; Algorithms; Bacteria; Biochemical; Biological Assay; Biological Models; Blood; Cardiovascular Diseases; Cause of Death; Cells; Characteristics; Chylomicrons; Complement; Conflict (Psychology); Crystallization; Data; Defect; Diabetes Mellitus; Disease; Endoplasmic Reticulum; Ensure; Enzymes; Eukaryota; Exhibits; Family; Family member; Foundations; Goals; Human; Hydrolase; Hydrolysis; Hypertriglyceridemia; In Vitro; Integral Membrane Protein; Lipase; Lipids; Location; Maps; Membrane; Membrane Proteins; Mentors; Methods; Modeling; Molecular Chaperones; Molecular Weight; Mycobacterium tuberculosis; Nonesterified Fatty Acids; Nutrient; Phase; Physiological; Physiology; Process; Protein Family; Proteins; Reporter; Resolution; Risk Factors; Role; Saccharomyces cerevisiae; Starvation; Structure; System; Triglycerides; Very low density lipoprotein; Work; genome sequencing; in vivo; lipid metabolism; lipoprotein lipase; member; novel; pathogen; pathogenic bacteria

Description: Hydrolysis of triacylglycerols by lipoprotein lipase (LPL) is essential for normal lipid metabolism. LPL does not fold and function properly without a newly identified, transmembrane protein, lipase maturation factor (LMF1). LMF1 is essentially uncharacterized; in fact it belongs to a family of proteins with a similar "domain of unknown function" (Duf1222). The major goal of this project is to investigate the interaction between LPL and LMF1. By analogy to lipases with structural characteristics similar to LPL, we propose that LMF1 acts as a "private foldase" for LPL. In Aim 1, I will use biochemical assays and assays in cells to determine why LPL needs folding assistance and how LMF1 helps LPL fold. Aim 2 consists of analysis of the membrane topology and structure of LMF1. Finally, Aim 3 focuses on the observation that many human pathogens have Duf1222 proteins; therefore I will use computational and biochemical approaches to determine if the Duf1222 protein represents one-half of a foldase-lipase pair. During the mentored phase, I will build on my preliminary observations to develop a model for LMF1 membrane topology. Additionally I will begin biochemical and structural analyses of the LMF-LPL interaction, and begin preliminary work on aim 3. For the independent phase, I will continue structural analysis of LMF1 and expand my biochemical analysis to Duf1222 proteins in other species to determine if they have important roles in lipid metabolism.

描述： 脂蛋白脂肪酶 (LPL) 水解三酰甘油对于正常脂质至关重要 代谢。如果没有新确定的、 跨膜蛋白、脂肪酶成熟因子（LMF1）。 LMF1本质上是 没有特征的；事实上，它属于具有相似“结构域”的蛋白质家族。 未知功能”（Duf1222）。该项目的主要目标是调查 LPL 和 LMF1 之间的相互作用。与具有结构特征的脂肪酶类比 与 LPL 类似，我们建议 LMF1 作为 LPL 的“私有折叠酶”。在目标 1 中，我将 使用生化分析和细胞分析来确定 LPL 需要折叠的原因 帮助以及 LMF1 如何帮助 LPL 弃牌。目标 2 包括膜分析 LMF1 的拓扑和结构。最后，目标 3 重点关注许多人的观察 人类病原体含有 Duf1222 蛋白；因此我将使用计算和 生化方法确定 Duf1222 蛋白是否代表一半 折叠酶-脂肪酶对。在指导阶段，我将在我的初步基础上 观察开发 LMF1 膜拓扑模型。另外我将开始 LMF-LPL 相互作用的生化和结构分析，并开始初步 致力于目标3。对于独立阶段，我将继续LMF1的结构分析 并将我的生化分析扩展到其他物种的 Duf1222 蛋白，以确定是否 它们在脂质代谢中具有重要作用。