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Functional characterization of a mitochondrial orphan enzyme

线粒体孤儿酶的功能表征

基本信息

批准号：
9120683
负责人：
Hongying Shen
金额：
$ 5.46万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2017-07-06
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9120683
关键词：
ATP Synthesis Pathway Acetyl Coenzyme A Amino Acids Anemia Biochemical Biochemistry Biological Assay CRISPR/Cas technology Carbon Cell Line Cells Cellular biology Cholesterol Citrates Citric Acid Cycle Coenzyme A Complement Complex Cultured Cells Detection Disease Enzymes Epitopes Exhibits Genes Genetic Genome engineering Genomics Goals Health Heart Heart Diseases Housing Human Human Activities Human Genetics Human Genome In Vitro Knock-out Label Laboratories Light Link Lipids Lyase Malates Mammals Mass Spectrum Analysis Mediating Metabolic Metabolic Pathway Metabolism Methionine Methods Methylmalonic Acid Methylmalonyl-CoA Mutase Mitochondria Mitochondrial Matrix Molecular Mutase Neurologic Dysfunctions Organelles Orphan Pathway interactions Physiological Physiology Play Population Process Propionic Acids Proteins Proteome Reaction Recombinants Recycling Regulation Research Role Techniques Testing Transcript Variant Vitamin B 12 Western Blotting Work base cell growth cobamamide cofactor genome wide association study glyoxylate human disease in vivo inhibitor/antagonist insight knock-down loss of function loss of function mutation lymphoblast metabolomics mitochondrial metabolism novel oxidation public health relevance skills small molecule thioester trait transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of the proposal is to decipher the function of mitochondrial orphan enzymes and their regulations on unexplored metabolic pathways with potential implications for human diseases. Mitochondria are intracellular organelles that house not only the machinery for ATP synthesis and fuel oxidation, but also metabolic enzymes for synthesis of building blocks for cell growth. For instance, a critical mitochondrial enzyme methylmalonyl-CoA mutase (MUT) recycles branched carbon chains from amino acids and lipids into tricarboxylic acid cycle. This process is critically dependent on an essential coenzyme vitamin B12 (B12). This project focuses on the role played by CLYBL, a ubiquitously expressed mitochondrial enzyme of unknown function, in regulating mitochondrial B12 function. A loss-of-function variant of CLYBL has been linked to subclinical B12 deficiency by two recent human genome-wide association studies. This proposal builds on the computational genomics analysis performed in our laboratory, which suggested CLYBL lies at the heart of a novel mitochondrial metabolic pathway connecting to B12-dependent MUT function. The proposed research seeks to understand: (1) enzymatic activities of CLYBL; (2) the CLYBL-dependent regulation of mitochondrial B12 function; and (3) molecular mechanism underlying this regulation. These studies will take advantage of the laboratory's expertise in advanced mass spectrometry-based metabolomics, computational genomics and mitochondrial physiology, combined with my skills in biochemistry and cell biology, to decipher the physiological function of CLYBL. This study of CLYBL represents a great example of "reverse human genetics". This work promises to shed new insights on mitochondrial biochemistry with potential implications for human diseases.

描述（由申请人提供）：该提案的长期目标是破译线粒体孤儿酶的功能及其对未探索的代谢途径的调节，对人类疾病具有潜在影响。线粒体是细胞内的细胞器，其不仅容纳用于ATP合成和燃料氧化的机器，而且还容纳用于合成细胞生长的构件的代谢酶。例如，一种关键的线粒体酶甲基丙二酰辅酶A（MUT）将来自氨基酸和脂质的支链碳链分解成三羧酸循环。这一过程严重依赖于必需的辅酶维生素B12（B12）。该项目的重点是CLYBL，一种广泛表达的功能未知的线粒体酶，在调节线粒体B12功能中所起的作用。最近的两项人类全基因组关联研究已将CLYBL的功能丧失变体与亚临床B12缺乏症联系起来。该提议建立在我们实验室进行的计算基因组学分析的基础上，该分析表明CLYBL位于连接B12依赖性MUT功能的新型线粒体代谢途径的核心。拟议的研究旨在了解：（1）CLYBL的酶活性;（2）线粒体B12功能的CLYBL依赖性调节;以及（3）这种调节的分子机制。这些研究将利用实验室在先进的基于质谱的代谢组学，计算基因组学和线粒体生理学方面的专业知识，结合我在生物化学和细胞生物学方面的技能，来破译CLYBL的生理功能。CLYBL的这项研究代表了“反向人类遗传学”的一个很好的例子。这项工作有望揭示线粒体生物化学的新见解，对人类疾病具有潜在的影响。