Strategie zur Identifizierung zyklischer Peptide als selektive Inhibitoren von RNA-Protein-Wechselwirkungen zur antibakteriellen Therapie

鉴定环肽作为抗菌治疗中 RNA-蛋白质相互作用的选择性抑制剂的策略

• 批准号: 211611392
• 负责人: Dr. Sabine Schneider
• 金额: --
• 依托单位: Department Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/211611392?language=en
• 关键词: Strategie; zur; Identifizierung; zyklischer; Peptide

Structured RNA-elements play an important role in the gene regulation of viruses, bacteria as well as eukrayotes. Bacteria use so called riboswitches in the mRNA to link gene expression to their metabolism. Binding of ligands to the riboswitch induces conformational changes resulting in the inhibition of transcription or translation of the respect gene. Cyclic peptides (ZP) belong to the most effective, high-affinity drug-like agents, which are extremely stable, flexible, variable and exhibit a wide range of biological activities. Recently methods were developed to generate libraries of genetically encoded and expressed cyclopeptides, which allow the direct identification of the active member when combined with an in vivo screening assay. Aim of the here proposed project is to establish an in vivo screening assay to identify ZPs from an expressed library, specifically binding to structured and conserved RNA elements in bacterial mRNAs in order to inhibit growth of pathogenic bacteria. These ZP-RNA interactions will be biochemical, biophysical and structural characterised and their conformational states theoretically and biophysically analysed. Based on these results, chemical modifications influencing the dynamics of the ZPs will be introduced, resulting in an increased binding probability and affinity to the target RNA.

结构化 RNA 元件在病毒、细菌和真核生物的基因调控中发挥着重要作用。细菌利用 mRNA 中所谓的核糖开关将基因表达与其代谢联系起来。配体与核糖开关的结合诱导构象变化，从而抑制相关基因的转录或翻译。环肽（ZP）属于最有效、高亲和力的类药物制剂，具有极其稳定、灵活、可变且表现出广泛的生物活性。最近开发了生成基因编码和表达环肽文库的方法，当与体内筛选测定相结合时，可以直接鉴定活性成员。这里提出的项目的目的是建立一种体内筛选方法，从表达文库中识别 ZP，特异性结合细菌 mRNA 中的结构化和保守的 RNA 元件，以抑制病原菌的生长。这些 ZP-RNA 相互作用将被进行生物化学、生物物理和结构表征，并从理论上和生物物理上分析它们的构象状态。基于这些结果，将引入影响 ZP 动力学的化学修饰，从而增加与目标 RNA 的结合概率和亲和力。