TRANSCARBOXYLASE, A 12MDA MULTIENZYME COMPLEX

转羧酶，一种 12MDA 多酶复合物

• 批准号: 7726278
• 负责人: VIVIEN YEE
• 金额: $ 0.58万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726278
• 关键词: Active Sites; Biological Models; Biotin; Catalysis; Complex; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Funding; Goals; Grant; Human; Institution; Ligands; Metabolic; Metabolic Diseases; Methylmalonyl-CoA carboxyltransferase; Multienzyme Complexes; Mutation; Publishing; Research; Research Personnel; Resources; Source; Structure; United States National Institutes of Health; dalton; design; enzyme mechanism; insight; polypeptide; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of the proposed research is to determine crystal structures of the transcarboxylase (TC) multienzyme complex. TC is a 1.2 million dalton biotin-dependent bacterial metabolic enzyme that has long served as a model system for human biotin-dependent metabolic enzymes; none of the human or mammalian biotin-dependent enzymes have been crystallized. Crystal structures of TC will provide insight into mechanisms of assembly and catalysis for biotin-dependent metabolic enzymes. Intact 26S TC contains 30 polypeptide chains of three types: the catalytic hexameric 12S, the catalytic dimeric 5S, and the biotinylated 1.3S which shuttles between the 12S and 5S active sites. The 26S TC can lose several 6S (5S+1.3S heterotetramers) subunits to give 18S, which is also enzymatically active. We have recently published crystal structures of the isolated 12S and 5S subunits, determined using synchrotron radiation. We propose to determine crystal structures of TC 6S, 18S, and 26S, as well as additional structures of isolated 12S and 5S, either as active site ligand complexes or mutations designed to probe enzyme mechanism or to reproduce the effect of human mutations identified in metabolic diseases.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 拟议的研究的目的是确定经羧化酶（TC）多烯酶复合物的晶体结构。 TC是一种120万个道尔顿生物素依赖性细菌代谢酶，长期以来一直是人类生物素依赖性代谢酶的模型系统。人类或哺乳动物依赖性酶均未结晶。 TC的晶体结构将深入了解依赖生物素的代谢酶的组装机理和催化。完整的26S TC包含三种类型的30种多肽链：催化六聚体12s，催化二聚体5s和生物素化的1.3s，它们在12s和5s活性位点之间穿梭。 26S TC可能会失去几个6s（5s+1.3s异驱动器）亚基以产生18s，这也具有酶活性。我们最近发表了使用同步加速器辐射确定的分离的12S和5S亚基的晶体结构。我们建议确定TC 6S，18S和26S的晶体结构，以及分离的12s和5s的其他结构，即作为活性位点配体复合物或旨在探测酶机制的突变，或重现在代谢疾病中鉴定出的人类突变的作用。