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（申请人提供）：描述：脂蛋白脂肪酶（LPL）对三酰基甘油的水解对于正常脂质代谢至关重要。如果没有新鉴定的跨膜蛋白，脂肪酶成熟因子（LMF1），LPL无法折叠并正常运行。 LMF1本质上是未表征的；实际上，它属于具有类似“未知功能领域”的蛋白质家族（DUF1222）。该项目的主要目标是研究LPL和LMF1之间的相互作用。类似于与LPL相似的结构特征的脂肪酶，我们建议LMF1充当LPL的“私有折叠酶”。在AIM 1中，我将在细胞中使用生化测定和测定，以确定LPL为什么需要折叠辅助以及LMF1如何帮助LPL折叠。 AIM 2包括对LMF1的膜拓扑和结构的分析。最后，AIM 3的重点是许多人类病原体具有DUF1222蛋白质的观察。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，我将基于我的初步观察，以开发LMF1膜拓扑模型。此外，我将开始对LMF-LPL相互作用的生化和结构分析，并开始在AIM 3上进行初步工作。对于独立阶段，我将继续对LMF1进行结构性分析，并将我的生化分析扩展到其他物种的DUF1222蛋白质，以确定它们在脂质代理中是否具有重要角色。 公共卫生相关性：相关性心血管疾病是发达国家的主要死亡原因，升高的血液三酰基甘油是危险因素。 LPL缺乏会导致血液三酰基甘油升高，干扰整体脂质代谢。 LMF1通过较知的过程促进LPL活性，阐明该机制将为开始改善LPL缺乏症的原因提供基础。