Mechanistic studies of thioether crosslink formation in peptides

肽中硫醚交联形成的机理研究

• 批准号: 9169902
• 负责人: VAHE BANDARIAN
• 金额: $ 28.7万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9169902
• 关键词: Active Sites; Address; Adoption; Anabolism; Anti-Bacterial Agents; Antineoplastic Agents; Antiviral Agents; Area; Binding; Biochemical; Carbon; Catalysis; Cells; Chemistry; Cleaved cell; Clinic; Cysteine; Electron Spin Resonance Spectroscopy; Electrons; Engineering; Enzymatic Biochemistry; Enzymes; Family; Future; Goals; Housing; Label; Lead; Life; Mediating; Methionine; Methods; Modification; Molecular; Mutation; Natural Products; Outcome; Oxidation-Reduction; Pathway interactions; Peptide Synthesis; Peptides; Play; Post-Translational Protein Processing; Property; Protein Family; Publishing; Reaction; Reporting; Research; Roentgen Rays; Role; Site; Structure; Structure-Activity Relationship; Sulfur; System; Therapeutic Agents; Therapeutic Uses; Time; Variant; base; cofactor; crosslink; enzyme mechanism; insight; killings; member; novel; oxidation; peptide synthase; polypeptide; quorum sensing; reconstitution; spectroscopic survey; thioether; tool

PROJECT SUMMARY All domains of life produce peptidic natural products that play roles as diverse as quorum sensing molecules and redox cofactors. The ability of these peptides to function as antibacterial, antiviral, or anticancer agents makes understanding their biosynthesis an important area of contemporary research. Peptide-based secondary metabolites are produced by distinct biosynthetic pathways that differ in whether the peptide is synthesized by the action of non-ribosomal peptide synthetases (NRPS), or ribosomally produced from a genomically encoded orf. These peptides are often extensively modified. While in the NRPS systems many of the modifications occur concurrently with peptide synthesis, the ribosomally encoded peptides undergo posttranslational modification. These so-called ribosomally encoded posttranslationally modified polypeptides (RiPP) are a new emerging class of polypeptides that have extensive modifications that are introduced by mechansims that are mostly poorly understood. This application will focus on enzymes introduce sulfur-to-alpha carbon thioether crosslinks into ribosomally encoded peptides to produce sactipeptides. These linkages are distinct from the well-studied lanthipeptide, where thioether crosslinks form between a Cys residue and a dehydrated Thr/Ser. The sactipeptide maturases are members of the radical SAM family of enzymes and catalyze thioether crosslinks by a radical- mediated reaction that is not understood. While The biochemical, spectroscopic, and structural studies in this application will lead to a mechanistic paradigm for this important class of enzymes. While sactipeptides themselves have various therapeutic uses, understanding the substrate profiles and mechanisms of these enzymes would provide a tool for synthesis of crosslinked peptides, which are increasing finding utility in the clinic.

项目摘要 生命的所有领域都会产生肽天然产品，它们发挥着像法定人数一样多样化的角色 感应分子和氧化还原辅助因子。这些肽起作用的能力 抗菌，抗病毒或抗癌剂使其了解其生物合成 当代研究的重要领域。基于肽的二级代谢产物是 由不同的生物合成途径产生的，在肽是否为 通过非核糖体肽合成酶（NRP）或核糖体的作用合成 由基因组编码的ORF产生。这些肽通常是广泛的 修改的。而在NRPS系统中，许多修改同时进行 肽合成，核糖体编码的肽经历翻译后发生 修改。这些所谓的核糖体编码后修饰 多肽（RIPP）是一类新的多肽，具有广泛的多肽 机器人引入的修改大多是理理的。 该应用将着重于酶引入硫至α碳硫纤维 交叉链接进入核糖体编码的肽以产生乙酰肽。这些联系 与良好的灯笼二肽不同，其中硫醚交联之间形成 cys残留物和脱水的thr/ser。乙酰肽成熟酶是 激进的SAM家族的酶家族，并通过自由基催化硫醚交联 介导的反应尚不清楚。而生化，光谱和 该应用中的结构研究将导致这一重要的机械范式 酶类。乙酰肽本身具有各种治疗用途，但 了解这些酶的底物概况和机制将提供 合成交联肽的工具，这些肽正在增加诊所中发现实用性。