Discovery and characterization of biocatalysts for ribosomally encoded alpha-N-methylated peptide natural products

核糖体编码的α-N-甲基化肽天然产物生物催化剂的发现和表征

• 批准号: 9980950
• 负责人: Michael F Freeman
• 金额: $ 35.98万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9980950
• 关键词: Affinity; Amides; Antibiotics; Antineoplastic Agents; Antiviral Agents; Binding; Biological; Biological Models; Cell Membrane Permeability; Chemicals; Cyclosporine; Drug Kinetics; Family; Family member; Goals; Immunosuppressive Agents; In Vitro; Kinetics; Measurement; Methods; Methylation; Natural Products; Pathway interactions; Penicillins; Peptides; Periodicity; Process; Production; Research; Resistance; Ribosomes; Role; Route; Source; Structure; Surface; Vertebral column; Vision; Work; catalyst; flexibility; fungus; improved; peptide drug; protein protein interaction; screening

PROJECT SUMMARY / ABSTRACT Since the discovery of penicillin in the 1920s, bioactive peptide natural products have been used as antibiotic, antiviral, immunosuppressive, and anti-cancer agents. Many of these bioactive peptides harbor backbone α-N-methylations and/or macrocyclizations, since these tailorings significantly improve peptide pharmacokinetics. As seen in the blockbuster immunosuppressant cyclosporin A, α-N-methylated peptides have increased structural rigidity, proteolytic resistance, and membrane permeability. Additionally, cyclic α-N- methylated peptides are able to bind large, flat surfaces with high affinity, making them attractive targets for disrupting protein-protein interactions. Despite these advantages, inefficient synthetic and in vitro processes hinder the production, screening, and optimization of α-N-methylated peptides. In addition, natural sources of amide backbone-methylated peptides were thought to be completely limited to inflexible nonribosomal peptide biosynthetic pathways. The goal of this work is to tease out the mechanistic and structural constraints of α-N- methylating biocatalysts within our newly discovered ribosomally encoded peptide natural product family called the borosins. Our first objective identifies a potent bioactive model system encoded in a basidiomycete fungus to tease out the rules and limitations of borosin α-N-methylation, N-to-C macrocyclization, and bioactivity. Our second objective focuses on structurally distinct borosin family members, where we will perform detailed kinetic measurements to tease out the mechanism for chemically challenging α-N-methylation. We also outline our longer-term visions to develop methods for studying peptide natural products in basidiomycete fungi as well as our efforts to tease out the potential biological roles of borosin metabolites. This research will create a diverse toolbox of flexible and efficient catalysts for the biological production, screening, and optimization of genetically templated bioactive α-N-methylated peptides.

项目摘要 /摘要 自1920年代发现青霉素以来，生物活性肽天然产品已被用作 抗生素，抗病毒，免疫抑制和抗癌药。其中许多生物活性胡椒港港口 骨干α-n-甲基化和/或大环化，因为这些调整可显着改善肽 药代动力学。如在重击中的免疫抑制剂环孢菌素A中所见，α-N-甲基化肽具有 结构刚度增加，蛋白水解耐药性和膜渗透性。此外，环状α-n- 甲基化的胡椒体能够结合具有高亲和力的较大的平坦表面，使其成为吸引人的目标 破坏蛋白质 - 蛋白质相互作用。尽管有这些优势，效率低下的合成和体外过程 阻碍了α-N-甲基化肽的生产，筛选和优化。此外，自然来源 酰胺骨架 - 甲基化的辣椒被认为完全局限于僵化的非透白色辣椒 生物合成途径。这项工作的目的是取消α-n-的机械和结构约束 我们新发现的核糖体编码肽天然产物家族中的甲基化生物催化剂称为 硼素。我们的第一个目标确定了碱性真菌中编码的潜在生物活性模型系统 阐明硼丝α-n-甲基化，n-to-c大环化和生物活性的规则和局限性。我们的 第二个目标侧重于结构上不同的硼醇家庭成员，我们将在其中执行详细的动力学 测量以取消化学具有挑战性的α-N-甲基化机制。我们也概述了我们的 长期的视觉开发用于研究碱性真菌中肽天然产物的方法以及 我们努力阐明硼醇代谢产物的潜在生物学作用。这项研究将创建潜水员 用于生物产生，筛选和优化的柔性和有效催化剂的工具箱 模板活化的生物活性α-N-甲基化肽。