Diphthamide biosynthesis

二邻苯二甲酰胺生物合成

• 批准号: 9032502
• 负责人: Hening Lin
• 金额: $ 29.85万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9032502
• 关键词: Address; Anabolism; Archaea; Bacterial Toxins; Biological Process; Cells; Chemistry; Cleaved cell; Crystallography; Diphtheria Toxin; Electron Spin Resonance Spectroscopy; Electron Transport; Enzymatic Biochemistry; Enzymes; Eubacterium; Eukaryota; Eukaryotic Cell; Generations; Genes; Genetic study; Goals; Grant; Health; Human; Hydrogen; In Vitro; Interdisciplinary Study; Lead; Ligase; Malignant Neoplasms; Mediating; Modification; Mutagenesis; Pathway interactions; Peptide Elongation Factor 2; Post-Translational Protein Processing; Protein Biosynthesis; Proteins; Pseudomonas aeruginosa toxA protein; Pyrococcus horikoshii; Reaction; Recombinants; Role; Techniques; Testing; Time; Tumor Suppressor Genes; United States National Institutes of Health; Yeasts; adapter protein; amidation; analog; design; in vivo; insight; interest; novel; prevent; protein complex; protein protein interaction; reconstitution; research study; spectroscopic survey; tumor; tumorigenesis

DESCRIPTION (provided by applicant): This A1 competitive renewal of GM088276 is a multidisciplinary collaboration to investigate the biosynthesis of diphthamide, a unique protein posttranslational modification that occurs on archaeal and eukaryotic translation elongation factor 2. This modification has been known for over 30 years and is the target of several bacterial toxins, including diphtheria toxin. However, the biosynthesis and biological function are still poorly understood. Previous studies by others suggest that there are five proteins (Dph1-5) required for the first two steps of the biosynthesis, while no proteins were identified for the thid (and last) amidation step. Interestingly, deletion of several of the biosynthesis genes is found in tumors. With previous grant support, we made a number of important findings, including the discovery of a novel radical SAM enzyme for the first step of diphthamide biosynthesis and the identification of two new proteins required for the last step of the biosynthesis. This renewal wil build on these finding to further understand the chemistry and enzymology of diphthamide biosynthesis and to elucidate the complete biosynthetic pathway for the first time. The interesting Fe-S enzyme chemistry that will be elucidated in this proposal will significantly expand the chemistry scope and mechanistic understanding of Fe-S enzymes. The understanding of diphthamide biosynthesis will help to understand the biological functions of diphthamide and why deletion of diphthamide biosynthesis genes promotes tumorigenesis.

描述（由申请人提供）：GM088276 的 A1 竞争性更新是一项多学科合作，旨在研究二苯甲酰胺的生物合成，二苯甲酰胺是一种发生在古菌和真核翻译延伸因子 2 上的独特蛋白质翻译后修饰。这种修饰已为人所知 30 多年，是包括白喉毒素在内的多种细菌毒素的目标。但其生物合成和生物学功能 仍然知之甚少。其他人之前的研究表明，生物合成的前两个步骤需要五种蛋白质 (Dph1-5)，而第三个（也是最后一个）酰胺化步骤没有发现任何蛋白质。有趣的是，发现几个生物合成基因的缺失 肿瘤。在之前的资助支持下，我们取得了许多重要发现，包括发现了一种用于二苯胺生物合成第一步的新型自由基 SAM 酶，以及鉴定了生物合成最后一步所需的两种新蛋白质。此次更新将建立在这些发现的基础上，进一步了解二邻苯二甲酰胺生物合成的化学和酶学，并首次阐明完整的生物合成途径。本提案中将阐明的有趣的 Fe-S 酶化学将显着扩展 Fe-S 酶的化学范围和机理理解。对二邻苯二甲酰胺生物合成的了解将有助于理解二邻苯二甲酰胺的生物学功能以及为什么二邻苯二甲酰胺生物合成基因的缺失会促进肿瘤发生。