Structure and regulation of lipid metabolism and transport

脂质代谢和运输的结构和调节

基本信息

批准号：
10622673
负责人：
Michael Virgil Airola
金额：
$ 43.95万
依托单位：
STATE UNIVERSITY NEW YORK STONY BROOK
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-08-01 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10622673
关键词：
Anabolism Binding Biochemical Biological Biological Assay Cardiovascular Diseases Carrier Proteins Cell Proliferation Cell membrane Complement Complex Deuterium Diabetes Mellitus Diglycerides Enzymes Fatty Acids Fatty acid glycerol esters Foundations Funding Goals Heart Diseases Hydrogen Hydrolysis Ions Knowledge Lecithin Link Lipid Biochemistry Lipids Lipoproteins Malignant Neoplasms Mammalian Cell Mass Spectrum Analysis Mediating Membrane Metabolism Molecular Molecular Conformation Molecular Mechanisms of Action Nuclear Envelope Obesity Phosphatidate Phosphatase Phosphatidic Acid Phosphatidylinositols Phospholipase D Phospholipids Physiological Play Post-Translational Protein Processing Process Protein Dephosphorylation Protein phosphatase Proteins Regulation Role Second Messenger Systems Site Stimulus Structure Substrate Specificity Triglycerides Vision Disorders Work cell motility extracellular human disease improved insight insulin sensitivity interest lipid metabolism lipid transfer protein lipid transport lipine pharmacologic protein function receptor response therapeutic target trafficking

项目摘要

PROJECT SUMMARY: We are interested in understanding how lipid-metabolizing enzymes and lipid transport proteins function and are regulated at the molecular and structural level. Previously, we focused on two enzymes in phosphatidic acid (PA) metabolism: lipin and phospholipase D (PLD). Lipins are PA phosphatases that dephosphorylate PA to generate diacylglycerol, which is the penultimate step in triglyceride biosynthesis. Lipins regulate phospholipid and lipoprotein synthesis, fat storage as triglycerides, fatty acid synthesis, and insulin sensitivity with implications to obesity, diabetes, and cardiovascular disease. PLDs hydrolyze phosphatidylcholine to produce the lipid second messenger PA in response to extracellular stimuli. Receptor- mediated activation of PLD regulates vesicular trafficking, cell proliferation, and cell migration, which has established them as therapeutic targets for cancer. Despite extensive studies on these highly regulated, multi- domain enzymes, significant gaps remain in our knowledge of their molecular mechanisms of action. During the past funding period, we determined the first structures of lipin and PLD, identified a new type of membrane- binding domain in lipins, and provided insight into PLD activation by lipids and protein effectors. These discoveries provide an excellent foundation for this application, where our main goals include 1) continuing to understand how PLD is activated by protein effectors and lipids, and 2) examining the molecular mechanisms regulating lipin PA phosphatase activity and PA substrate specificity. We will also 3) biochemically and structurally characterize the nuclear envelope protein phosphatase complex CTDNEP1-NEP1R1, which dephosphorylates lipin to control lipin PA phosphatase activity; and 4) study the structure and function of Nir lipid transfer proteins, which transfer PA and phosphatidylinositol between membrane contact sites to replenish pools of plasma membrane phosphoinositides after hydrolysis by PLC. Structural studies will be complemented by an array of lipid biochemistry and lipid-protein interaction assays that we are well versed in, hydrogen-deuterium exchange mass spectrometry, and cellular studies in mammalian cells. A network of collaborators who are leaders in their respective fields supports these studies. We aim to answer several major questions: (1) How does the structure of lipid-modifying enzymes allow them to recognize their lipid substrates and interact with the membrane? (2) What role do structurally and/or functionally uncharacterized domains play in the action of these proteins? (3) How are these enzymes/proteins regulated? Specifically, are they autoinhibited? how do protein effectors, lipids, ions, and post-translational modifications regulate activity? and what conformational changes occur during activation? Overall, this work will improve our understanding of biological mechanisms and provide information on lipid-protein interactions of physiological and pharmacological significance.

项目摘要：我们有兴趣了解脂质 - 代谢酶和脂质运输如何蛋白质功能，并在分子和结构水平上进行调节。以前，我们专注于两个酶在磷脂酸（PA）代谢中：脂蛋白和磷脂酶D（PLD）。脂肪是PA磷酸酶去磷酸酯PA产生二酰基甘油，这是甘油三酸酯生物合成的倒数第二步。 Lipins 调节磷脂和脂蛋白合成，脂肪储存为甘油三酸酯，脂肪酸合成和胰岛素对肥胖，糖尿病和心血管疾病的敏感性。 PLD水解磷脂酰胆碱可用于响应细胞外刺激而产生脂质第二信使PA。受体 PLD的介导的激活调节囊泡运输，细胞增殖和细胞迁移，这具有将它们确定为癌症的治疗靶标。尽管对这些高度调节的多种研究领域酶，我们对它们的分子作用机理的了解仍然存在很大的差距。在过去的资金期，我们确定了Lipin和PLD的第一个结构，确定了一种新型的膜 - 脂肪中的结合结构域，并提供了脂质和蛋白质效应子的PLD激活的见解。这些发现为此应用提供了绝佳的基础，我们的主要目标包括1）继续了解如何通过蛋白质效应子和脂质激活PLD，以及2）检查分子机制调节脂蛋白PA磷酸酶活性和PA底物特异性。我们还将3）生化和在结构上表征了核包膜蛋白磷酸酶复合物CTDNEP1-NEP1R1，该蛋白去磷酸化Lipin以控制脂肪蛋白PA磷酸酶活性； 4）研究NIR脂质的结构和功能转移蛋白质，将PA和磷脂酰肌醇转移到膜接触位点之间以补充池通过PLC水解后质膜磷酸肌醇的质膜。结构研究将由脂质生物化学和脂质 - 蛋白质相互作用测定阵列，我们精通氢螺旋体交换质谱和哺乳动物细胞中的细胞研究。一个合作者网络各自领域的领导者支持这些研究。我们的目标是回答几个主要问题：（1）脂质修饰酶的结构是否使它们能够识别其脂质底物并与膜？（2）在结构上和/或功能上未表征的域在这些作用中起什么作用蛋白质？（3）如何调节这些酶/蛋白质？具体来说，它们是自身抑制的吗？蛋白质如何效应子，脂质，离子和翻译后修饰调节活性？以及什么构象变化在激活过程中发生？总体而言，这项工作将提高我们对生物机制的理解，并提供有关生理和药理意义的脂质蛋白相互作用的信息。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Targeting Sterylglucosidase A to Treat Aspergillus fumigatus Infections.

DOI：
10.1128/mbio.00339-23
发表时间：
2023-04-25
期刊：
mBio
影响因子：
6.4
作者：
通讯作者：

Publisher Correction: Crystal structure of a lipin/Pah phosphatidic acid phosphatase.

出版商更正：脂质/Pah 磷脂酸磷酸酶的晶体结构。

DOI：
10.1038/s41467-020-15509-0
发表时间：
2020
期刊：
Nature communications
影响因子：
16.6
作者：
Khayyo,ValerieI;Hoffmann,ReeceM;Wang,Huan;Bell,JustinA;Burke,JohnE;Reue,Karen;Airola,MichaelV
通讯作者：
Airola,MichaelV

The middle lipin domain adopts a membrane-binding dimeric protein fold.

DOI：
10.1038/s41467-021-24929-5
发表时间：
2021-08-05
期刊：
Nature communications
影响因子：
16.6
作者：
Gu W;Gao S;Wang H;Fleming KD;Hoffmann RM;Yang JW;Patel NM;Choi YM;Burke JE;Reue K;Airola MV
通讯作者：
Airola MV

Structure and mechanism of the human CTDNEP1-NEP1R1 membrane protein phosphatase complex necessary to maintain ER membrane morphology.

维持内质网膜形态所必需的人 CTDNEP1-NEP1R1 膜蛋白磷酸酶复合物的结构和机制。