The effect of mitochondrial CoA degradation on glucose and fatty acid metabolism

线粒体CoA降解对葡萄糖和脂肪酸代谢的影响

• 批准号: 9891848
• 负责人: Evan W Kerr
• 金额: $ 3.92万
• 依托单位: WEST VIRGINIA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-17 至 2020-12-19
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9891848
• 关键词: ATP Synthesis Pathway; Biochemical; Biochemistry; Biological Assay; Blood Glucose; Brown Fat; Cell Fractionation; Cell Respiration; Cells; Coenzyme A; Collaborations; Coupled; Critical Thinking; Cytosol; Data; Dependovirus; Diabetes Mellitus; Disease; Drug or chemical Tissue Distribution; Enzymes; Equilibrium; Fasting; Fatty Acids; Gluconeogenesis; Glucose; Heart; Hepatic; Hepatocyte; Hyperglycemia; Immunofluorescence Immunologic; Ketone Bodies; Kidney; Knowledge; Lead; Link; Lipids; Liver; Measures; Mediating; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Mitochondria; Mus; Muscle; NADH dehydrogenase (ubiquinone); Neurodegenerative Disorders; Non-Insulin-Dependent Diabetes Mellitus; Nutritional; Oral; Pathway interactions; Phenotype; Physiological; Play; Proteins; Regulation; Research; Research Personnel; Research Training; Reverse Transcriptase Polymerase Chain Reaction; Role; Scientist; Sequence Homology; Skeletal Muscle; Substrate Specificity; Techniques; Testing; Tissues; Training; Universities; West Virginia; Western Blotting; Work; acyl group; cancer type; cofactor; collaborative environment; diabetic; fatty acid metabolism; fatty acid oxidation; glucose metabolism; glucose production; insight; ketogenesis; knowledge base; member; metabolomics; mitochondrial metabolism; novel; nudix hydrolase; overexpression; oxidation; peroxisome; phosphodiester; research facility; response; skills; thioester

PROJECT SUMMARY/ABSTRACT Mitochondrial metabolism is dependent on coenzyme A (CoA) to support metabolic processes like fatty acid oxidation, ketogenesis, and gluconeogenesis. Accumulation of CoA results in increased fatty acid oxidation, which drives gluconeogenesis in diabetes. Abnormally high CoA levels in skeletal muscle blunt mitochondrial ATP synthesis by decreasing the activity of complex I of the electron transport chain. Despite this evidence linking increased CoA to metabolic dysregulation, the mechanisms regulating CoA levels within the mitochondria are currently unknown. Nudt8 is an uncharacterized member of the Nudix hydrolase superfamily. This protein has been annotated as a potential CoA-degrading enzyme due to its high sequence homology to Nudt7, a previously characterized CoA diphosphohydrolase that resides in the peroxisomes. Our preliminary data show that Nudt8 specifically degrades free CoA and CoA thioesters. Additionally, Nudt8 localizes to the mitochondria and is expressed in highly oxidative tissues such as the heart, kidney, liver, brown adipose tissue, and muscle. The CoA-degrading activity of Nudt8, coupled with its localization to the mitochondria, suggests a role for this enzyme in the regulation of mitochondrial CoA levels and thus, metabolism. The proposed research will utilize adeno-associated virus to achieve over-expression of Nudt8 in the liver, followed by analysis of its effects on mitochondrial CoA pool size and composition, and on the mitochondrial metabolism of fatty acids and glucose. Additionally, mechanisms regulating Nudt8 expression and activity will be elucidated. The proposed research will be completed with the support of the excellent research facilities available to the applicant and in collaboration with other investigators within and outside the Department of Biochemistry at West Virginia University. The highly collaborative environment, combined with a personalized training plan, will allow the applicant to acquire skills in a broad range of techniques and to develop critical thinking skills, both of which are essential for a trainee to develop into a successful scientist. The applicant will participate in rigorous course work and oral presentations; these practices will provide an extensive knowledge base in metabolism and enhance the ability of the applicant to communicate his scientific ideas. Overall, the proposed combination of research and training plans will produce a well-rounded scientist as well as valuable insight into the regulation of the mitochondrial CoA pool and potential of targeting Nudt8 to correct the dysregulated mitochondrial metabolism that underlies a variety of metabolic diseases.

项目摘要/摘要 线粒体代谢取决于辅酶A（COA）以支持脂肪酸等新陈代谢过程 氧化，生酮发生和糖异生。 COA的积累导致脂肪酸氧化增加， 这驱动糖尿病中的糖异生。骨骼肌钝线线粒体异常高的COA水平 通过减少电子传输链复合物I的活性来合成ATP。尽管有这些证据 将增加的COA与代谢失调联系起来，该机制调节线粒体内的COA水平 目前未知。 NUDT8是Nudix水解酶超家族的未表征。该蛋白质 由于其与Nudt7的高序列同源，A 先前对过氧化物酶体中的COA双磷酸化酶进行了表征。我们的初步数据显示 NUDT8专门降低了免费的COA和COA Thioesters。此外，NUDT8本地化在线粒体上 并在高度氧化组织中表达，例如心脏，肾脏，肝脏，棕色脂肪组织和肌肉。 NUDT8的CoA降解活性以及其本地化与线粒体的本地化表明了这一点 线粒体COA水平的调节中的酶，因此是代谢。拟议的研究将利用 腺相关病毒以实现肝脏中NUDT8的过表达，然后分析其对 线粒体COA池的大小和组成，以及脂肪酸和葡萄糖的线粒体代谢。 另外，将阐明调节NUDT8表达和活性的机制。 拟议的研究将在可用的优秀研究设施的支持下完成 申请人并与West生物化学部内外的其他研究人员合作 弗吉尼亚大学。高度协作的环境以及一个个性化的培训计划，将允许 申请人在广泛的技术中获取技能并发展批判性思维技能，这两者都 对于学员发展成为成功的科学家至关重要。申请人将参加严格的课程 工作和口头演示；这些实践将在代谢和 增强申请人传达其科学思想的能力。总体而言，提议的组合 研究和培训计划将产生全面的科学家，并对该法规的宝贵见解 线粒体COA池的量和靶向NUDT8的潜力校正非调节线粒体的失调 代谢是各种新陈代谢疾病的基础。