A Universal Support for the Solid-Phase Synthesis of Peptidyl-tRNA Mimics.

IF 2.6 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
ChemBioChem Pub Date : 2024-10-28 DOI:10.1002/cbic.202400717
Julia Thaler, Christoph Mitteregger, Laurin Flemmich, Ronald Micura
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引用次数: 0

Abstract

Hydrolysis-resistant RNA-peptide conjugates that mimic peptidyl-tRNAs are often required for structural and functional studies of protein synthesis at the ribosome. These conjugates can be synthesized by solid-phase chemical synthesis, which allows maximum flexibility in both the peptide and RNA sequence. The commonly used strategy is based on (3'-N-aminoacyl)-3'-amino-3'-deoxyadenosine solid supports, which already contain the first C-terminal amino acid of the target peptidyl chain. This is a limitation in the sense that different individual supports must be synthesized for different C-terminal amino acids. In this study, we demonstrate a solution to this problem by introducing a novel universal support. The key is a free ribose 3'-NH2 group that can be coupled to any amino acid. This is made possible by a photocleavable ether moiety that links the ribose 2'-O to the support, thus avoiding the typical O-to-N migration that occurs when using 2'-O-acyl linked solid supports. Once assembled, the conjugate is readily cleaved by UV irradiation. The structural integrity of the obtained peptidyl-RNA conjugates was verified by mass spectrometry analysis. In conclusion, the new photocleavable solid support makes the synthesis of 3'-peptidyl tRNA mimics of different peptidyl chains significantly more efficient compared to the commonly used approaches.

肽基-tRNA 模拟物固相合成的通用支持。
在核糖体蛋白质合成的结构和功能研究中,经常需要能模拟肽基-tRNA 的抗水解 RNA 肽共轭物。这些共轭物可通过固相化学合成法合成,从而使肽和 RNA 序列具有最大的灵活性。常用的策略是基于(3'-N-氨基酰基)-3'-氨基-3'-脱氧腺苷固体支持物,这种支持物已经含有目标肽链的第一个 C 端氨基酸。这就限制了我们必须为不同的 C 端氨基酸合成不同的支持物。在本研究中,我们通过引入一种新型通用支持物,展示了解决这一问题的方法。其关键在于可与任何氨基酸偶联的自由核糖 3'-NH2 基团。这是通过一个可光裂解的醚基将核糖 2'-O 与支撑物连接起来实现的,从而避免了使用 2'-O-acyl 链接固体支撑物时典型的 O-N 迁移。一旦组装完成,共轭物很容易在紫外线照射下裂解。质谱分析验证了所获得的肽基-RNA 共轭物的结构完整性。总之,与常用的方法相比,新的可光裂解固体支持物使不同肽链的 3'-肽基 tRNA 模拟物的合成效率显著提高。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
ChemBioChem
ChemBioChem 生物-生化与分子生物学
CiteScore
6.10
自引率
3.10%
发文量
407
审稿时长
1 months
期刊介绍: ChemBioChem (Impact Factor 2018: 2.641) publishes important breakthroughs across all areas at the interface of chemistry and biology, including the fields of chemical biology, bioorganic chemistry, bioinorganic chemistry, synthetic biology, biocatalysis, bionanotechnology, and biomaterials. It is published on behalf of Chemistry Europe, an association of 16 European chemical societies, and supported by the Asian Chemical Editorial Society (ACES).
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