RNA-controlled synthesis of peptides via peptidyl transfer reactions with peptide-nucleic acid conugates

通过肽-核酸缀合物的肽基转移反应进行 RNA 控制的肽合成

基本信息

批准号：
225213878
负责人：
Professor Dr. Oliver Seitz
金额：
--
依托单位：
Institut für Chemie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2012
资助国家：
德国
起止时间：
2011-12-31 至 2015-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/225213878?language=en
关键词：
RNA controlled synthesis peptides via

项目摘要

It is the aim to develop a reaction, which proceeds only when a specific RNA sequence is present and leads to the formation of a biologically active compound. This is accomplished by means of a RNA-triggered peptidyl transfer reaction, which bears resemblance to a key step in ribosomal peptide synthesis. The reaction involves a donor peptide, which is connected via a thioester bond to a peptide nucleic acid (PNA) unit. A second peptide-PNA conjugate offers a reactive 1,2- or 1,3-aminothiol structure at the N-terminal end and serves as acceptor. The two conjugates bind adjacently to a complementary RNA template. This triggers the intramolecular transfer of the donor onto the acceptor peptide. A new peptide bond is formed in a native chemical ligation like reaction. The prerequisites for efficient peptidyl transfer are examined by varying the architecture of the RNA templates, the length of the donor and acceptor peptides as well as the structure of the thioesters involved. The RNA-triggered peptidyl transfer may provide new opportunities for the development of a) artificial feedback loops in synthetic biology and b) cell type specific, personalized bioactive compounds. It is the long-term objective to hijack cell endogenous RNA molecules and use the RNA sequence information for the programmed formation of compounds that modulate signal transduction e.g. in cancer cells. The RNA templates can act as catalysts which trigger the formation of many peptide molecules per template. The achievable turnover rate is an important reaction parameter given that the RNA target of interest may occur at low abundance. The in situ formed peptides will be used to inhibit Bcl-xL; a protein that prevents apoptosis in cancer cells. Further efforts are directed to the RNA-induced synthesis of a phosphopeptide that inhibits the Signal Transduction and Activator of Transcription 3 (Stat3). The ability to translate RNA information into protein inhibition will be assessed in protein binding assays which involve recombinant proteins and proteins in cell lysates.

它的目的是发展反应，仅当存在特定的RNA序列并导致生物活性化合物的形成时才进行。这是通过RNA触发的肽基转移反应来完成的，该反应与核糖体肽合成的关键步骤相似。该反应涉及一个供体肽，该肽通过硫酯键连接到肽核酸（PNA）单位。第二肽-PNA结合物在N末端提供反应性的1,2-或1,3-氨基硫醇结构，并作为受体。两个偶联物与互补的RNA模板相邻结合。这会触发供体的分子内转移到受体肽上。在天然化学连接（如反应）中形成了一种新的肽键。通过改变RNA模板的结构，供体和受体肽的长度以及所涉及的硫代植物的结构来检查有效肽基转移的先决条件。 RNA触发的肽基转移可能为开发a）合成生物学中的人工反馈回路提供新的机会，而b）特定于细胞类型的个性化生物活性化合物。这是劫持细胞内源性RNA分子的长期目标，并使用RNA序列信息来编程形成化合物，以调节信号转导，例如在癌细胞中。 RNA模板可以充当催化剂，该催化剂触发每个模板许多肽分子的形成。鉴于感兴趣的RNA目标可能发生在低丰度时，可实现的周转率是一个重要的反应参数。原位形成的肽将用于抑制Bcl-XL；一种阻止癌细胞凋亡的蛋白质。进一步的努力针对RNA诱导的磷酸肽的合成，该磷酸肽抑制了转录3的信号转导和激活剂（STAT3）。将RNA信息转化为蛋白质抑制的能力将在蛋白质结合测定中进行评估，蛋白质结合测定法涉及细胞裂解物中的重组蛋白和蛋白质。