Engineering of a Peptide a-N-Methyltransferase to Methylate Non-Proteinogenic Amino Acids
Engineering of a Peptide a-N-Methyltransferase to Methylate Non-Proteinogenic Amino Acids
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肽α-N-甲基转移酶的工程改造以甲基化非蛋白氨基酸
DOI：
10.1002/ange.202100818
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发表时间：
2021
影响因子：
--
通讯作者：
Song H
中科院分区：
文献类型：
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作者：
Song H
在多肽中引入α‐N‐甲基化的非蛋白氨基酸可以提高其生物活性、膜通透性和蛋白水解稳定性。这通常是通过组装预甲基化氨基酸在自然界和实验室中实现的。甲基化酰胺键的更有吸引力的途径是具有挑战性的。生物学已经进化出一种α - N -自甲基化酶OphMA，它作用于融合到其C端的肽的酰胺键。由于其底物的核糖体生物合成，该酶对具有非蛋白质原性氨基酸的肽的活性尚未得到解决。一种工程化的OphMA，内部介导的蛋白质连接和固相肽合成使我们能够在非天然酰胺的情况下证明酰胺键的甲基化。该方法可应用于治疗性多肽的生物技术生产。
Introduction of α‐N‐methylated non‐proteinogenic amino acids into peptides can improve their biological activities, membrane permeability and proteolytic stability. This is commonly achieved, in nature and in the lab, by assembling pre‐methylated amino acids. The more appealing route of methylating amide bonds is challenging. Biology has evolved an α‐N‐automethylating enzyme, OphMA, which acts on the amide bonds of peptides fused to its C‐terminus. Due to the ribosomal biosynthesis of its substrate, the activity of this enzyme towards peptides with non‐proteinogenic amino acids has not been addressed. An engineered OphMA, intein‐mediated protein ligation and solid‐phase peptide synthesis have allowed us to demonstrate the methylation of amide bonds in the context of non‐natural amides. This approach may have application in the biotechnological production of therapeutic peptides.