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: 7770199
• 负责人: Saskia Neher
• 金额: $ 10.26万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7770199
• 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; public health relevance

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: Relevance Cardiovascular disease is a leading cause of death in the developed world, and elevated blood triacylglycerol is a risk factor. LPL deficiency results in elevated blood triacylglycerol and disturbs overall lipid metabolism. LMF1 promotes LPL activity through a poorly understood process, and elucidation of this mechanism will provide a foundation to begin to ameliorate one cause of LPL deficiency.

描述（由申请人提供）： 描述：脂蛋白脂肪酶（LPL）对三酰甘油的水解对于正常的脂质代谢至关重要。如果没有新鉴定的跨膜蛋白脂肪酶成熟因子 (LMF1)，LPL 就无法正常折叠和发挥作用。 LMF1 本质上是未表征的；事实上，它属于具有类似“未知功能域”（Duf1222）的蛋白质家族。该项目的主要目标是研究 LPL 和 LMF1 之间的相互作用。通过类比结构特征与 LPL 相似的脂肪酶，我们提出 LMF1 作为 LPL 的“私有折叠酶”。在目标 1 中，我将使用生化测定和细胞测定来确定 LPL 为何需要折叠辅助以及 LMF1 如何帮助 LPL 折叠。目标 2 包括分析 LMF1 的膜拓扑和结构。最后，目标 3 重点观察许多人类病原体都含有 Duf1222 蛋白；因此，我将使用计算和生化方法来确定 Duf1222 蛋白是否代表折叠酶-脂肪酶对的一半。在指导阶段，我将根据我的初步观察开发 LMF1 膜拓扑模型。此外，我将开始对 LMF-LPL 相互作用进行生化和结构分析，并开始目标 3 的初步工作。对于独立阶段，我将继续对 LMF1 进行结构分析，并将我的生化分析扩展到其他物种的 Duf1222 蛋白，以确定它们是否在脂质代谢中具有重要作用。 公共卫生相关性： 相关性 心血管疾病是发达国家死亡的主要原因，而血液三酰甘油升高是一个危险因素。 LPL 缺乏会导致血液三酰甘油升高并扰乱整体脂质代谢。 LMF1 通过一种鲜为人知的过程促进 LPL 活性，阐明这一机制将为开始改善 LPL 缺乏的一个原因奠定基础。