N-acetylglutamate Synthase: Structure, Function & Defects

N-乙酰谷氨酸合成酶：结构、功能

• 批准号: 7809804
• 负责人: Mendel Tuchman
• 金额: $ 55.88万
• 依托单位: CHILDREN'S RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7809804
• 关键词: Acetyl Coenzyme A; Address; Affect; Amino Acid Substitution; Ammonia; Anabolism; Animal Model; Animals; Archives; Arginine; Bacteria; Be++ element; Behavior Therapy; Beryllium; Binding; Biochemical; Biochemistry; Biological; Biology; Birth; Brain; Breeding; CYP21A2 gene; Carbamyl Phosphate; Catalysis; Citrulline; Clinical; Clinical Trials; Communities; Country; Crystallization; DNA Footprint; Data; Defect; Development; Diagnosis; Discipline; Disease; ES Cell Line; Embryo; Engineering; Enzyme Inhibition; Enzymes; Escherichia coli; Evolution; Functional disorder; Funding; Gene Expression; Gene Proteins; Genes; Genetic Transcription; Genotype; Glutamates; Goals; Grant; Health; Hepatic; Homozygote; Hyperammonemia; Hypersensitivity; Inherited; Intestines; Investigation; Kidney; Knock-out; Knockout Mice; Knowledge; Letters; Ligands; Ligase; Liver; Liver diseases; Mammals; Messenger RNA; Metabolism; Methods; Mitochondrial Matrix; Modeling; Molecular; Mus; Mutation; N acetyl L glutamate; N-carbamylglutamate; Neisseria gonorrhoeae; Nitrogen; Occupations; Organ; Organism; Patients; Phenotype; Phosphotransferases; Phylogenetic Analysis; Physiological; Physiology; Plants; Play; Poison; Primer Extension; Production; Property; Proteins; Proteobacteria; Race; Recombinant Proteins; Recombinants; Recovery; Regulation; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Site; Small Intestines; Structure; System; Tetraodontidae; Time; Tissues; Toxic effect; Transcription Initiation Site; Transcriptional Regulation; Transgenic Mice; Transgenic Model; United States National Institutes of Health; Urea; Vertebrates; Withdrawal; X-Ray Crystallography; Xanthomonas; Xanthomonas campestris; Xylella; Zebrafish; analog; arginylglutamate; base; chromatin immunoprecipitation; cofactor; fungus; hepatic ureagenesis; in vivo; knock out mouse project; mouse argA protein; mouse model; mutant; nitrogen balance; nitrogen metabolism; novel; parent grant; parent project; postnatal; prenatal; protein K; protein folding; protein profiling; protein structure; public health relevance; repository; research study; response; structural biology; three dimensional structure; tool; urea cycle

DESCRIPTION (provided by applicant): This is a Competitive Revision Application in response to the Recovery Act Funds Notice Number NOT-OD-09- 058. The parent project over goal is to explore the biology, biochemistry and pathophysiology of N- acetylglutamate synthase (NAGS) is an enzyme that produces the cognate cofactor N-acetylglutamate (NAG), an essential allosteric activator in ureagenesis. This revision application adds two new aims to the parent project. 1. To develop a knockout mouse for NAGS deficiency, characterize its phenotype, and explore its use as a conditional hyperammonemia model. 2. To "isolate" and study the in vivo regulation of ureagenesis specifically at the level of NAGS/CPSI by comparing nitrogen metabolism in N-carbamylglutamate (NCG) treated koNAGS mouse vs. wild type littermates. In addition to these new aims to be completed in two years, this project will enhance in the long term two of the existing aims that study the arginine effects on NAGS function and the effect of naturally-occurring mutations in patients with NAGS deficiency. In this revised project, we will develop, study and make available to the research community a novel "titratable mouse model of hyperammonemia. This knockout NAGS mouse will be rescued with NCG and will develop hyperammonemia upon withdrawal of this cofactor analog. We will determine the in vivo differences between nitrogen balance and metabolism in the NAGS "regulation-deprived" koNAGS mice rescued with NCG vs. the naturally-regulated wild type littermates on and off NCG. These studies will use both gene expression and protein profiles methods and will allow for the first time to "isolate" in vivo the regulatory effects of NAGS on ureagenesis. This project will enhance the pace and quality of the parent grant by providing a new tool for in vivo investigations for our group and other investigators studying hyperammonemia. In addition, The contribution of this project to the economy is leveraged by making the koNGAS mouse model available to othr investigators across the country, enhancing their research and promoting new job creation. PUBLIC HEALTH RELEVANCE: This project is dedicated to the investigation of an important gene and protein (NAGS) that determined how much nitrogen we eliminate from our bodies. It is important to know this since one of the main problem in liver disease is the inability to eliminate toxic nitrogen (ammonia) which can poison the brain. We will study an engineered mouse that does not posses NAGS to allow us to better understand this system and how it is regulated. The results from this project could allow the development of new treatments for elevated ammonia levels to protect the brain from its toxic effects.

描述（由申请人提供）：这是一份竞争性修订申请，以响应恢复法案资金通知编号no - od -09- 058。N-乙酰谷氨酸合成酶（N -acetylglutamate synthase， NAGS）是一种产生同源辅酶N-乙酰谷氨酸（N -acetylglutamate， NAG）的酶，是尿素生成过程中必不可少的变构激活剂。此修订应用程序向父项目添加了两个新目标。1. 建立NAGS基因敲除小鼠，确定其表型，并探讨其作为条件性高氨血症模型的应用。2. 通过比较n -氨甲酰谷氨酸（N-carbamylglutamate， NCG）处理的koNAGS小鼠与野生型幼崽的氮代谢，“分离”并研究在NAGS/CPSI水平上对尿生成的体内特异性调控。除了这些将在两年内完成的新目标外，该项目还将长期加强现有的两个目标，即研究精氨酸对NAGS功能的影响以及自然发生的突变对NAGS缺乏症患者的影响。在这个修订后的项目中，我们将开发、研究并向研究界提供一种新的“可滴定的高氨血症小鼠模型”。这种敲除NAGS的小鼠将被NCG拯救，并在退出这种辅助因子类似物后出现高氨血症。我们将测定用NCG拯救的NAGS“缺乏调节”的koNAGS小鼠与自然调节的野生型幼崽在NCG和关闭NCG时体内氮平衡和代谢的差异。这些研究将同时使用基因表达和蛋白质谱方法，并将首次允许在体内“分离”NAGS对尿致变性的调节作用。该项目将通过为我们小组和其他研究高氨血症的研究人员提供体内研究的新工具，提高父母资助的速度和质量。此外，该项目对经济的贡献是通过将koNGAS小鼠模型提供给全国其他研究人员，加强他们的研究并促进新的就业机会。