Ribosomal Elongation of Cyclic γ-Amino Acids using a Reprogrammed Genetic Code
Ribosomal Elongation of Cyclic γ-Amino Acids using a Reprogrammed Genetic Code
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DOI：
10.1021/jacs.9b12280
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发表时间：
2020-03-18
影响因子：
15
通讯作者：
Suga, Hiroaki
中科院分区：
文献类型：
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作者：
Katoh, Takayuki;Suga, Hiroaki
由于γ -氨基酸在羧基上酯化后通常会进行快速的自环化，例如γ -氨基酰基- trna，据我们所知，还没有关于γ -氨基酸核糖体延伸的报道。为了避免这种自环化，我们利用环状γ -氨基酸，并证明了它们的延伸成肽链。虽然环γ -氨基酸的掺入本质上是缓慢的，但我们在这里表明，伸长因子P和工程化trna的结合提高了环γ -氨基酸的掺入效率。通过这种方法，在重编程的遗传密码下表达了不仅含有环γ -氨基酸，还含有d - α -、n -甲基α -和环β -氨基酸的硫醚大环肽。含有环γ -氨基酸的核糖体合成的大环肽文库应适用于体外筛选方法，如mRNA展示，以发现新的肽药物。
Because gamma-amino acids generally undergo rapid self-cyclization upon esterification on the carboxyl group, for example, gamma-aminoacyl-tRNA, there are no reports of the ribosomal elongation of gamma-amino acids to the best of our knowledge. To avoid such self-cyclization, we utilized cyclic gamma-amino acids and demonstrated their elongation into a peptide chain. Although the incorporation of the cyclic gamma-amino acids is intrinsically slow, we here show that the combination of elongation factor P and engineered tRNAs improves cyclic gamma-amino acid incorporation efficiency. Via this method, thioether-macrocyclic peptides containing not only cyclic gamma-amino acids but also D-alpha-, N-methyl-alpha-, and cyclic beta-amino acids were expressed under the reprogrammed genetic code. Ribosomally synthesized macrocyclic peptide libraries containing cyclic gamma-amino acids should be applicable to in vitro screening methodologies such as mRNA display for discovering novel peptide drugs.