Department of Chemistry and Pharmacy, Institute of Biochemistry, Corrensstrasse 36, 48149, Münster, Germany.
Cells in Motion Interfaculty Center, University of Münster, Germany.
Angew Chem Int Ed Engl. 2021 Jun 7;60(24):13280-13286. doi: 10.1002/anie.202100352. Epub 2021 May 6.
Eukaryotic mRNAs are emerging modalities for protein replacement therapy and vaccination. Their 5' cap is important for mRNA translation and immune response and can be naturally methylated at different positions by S-adenosyl-l-methionine (AdoMet)-dependent methyltransferases (MTases). We report on the cosubstrate scope of the MTase CAPAM responsible for methylation at the N -position of adenosine start nucleotides using synthetic AdoMet analogs. The chemo-enzymatic propargylation enabled production of site-specifically modified reporter-mRNAs. These cap-propargylated mRNAs were efficiently translated and showed ≈3-fold increased immune response in human cells. The same effects were observed when the receptor binding domain (RBD) of SARS-CoV-2-a currently tested epitope for mRNA vaccination-was used. Site-specific chemo-enzymatic modification of eukaryotic mRNA may thus be a suitable strategy to modulate translation and immune response of mRNAs for future therapeutic applications.
真核生物 mRNA 正成为蛋白质替代疗法和疫苗接种的新兴模式。其 5' 帽结构对于 mRNA 翻译和免疫反应非常重要，并且可以被 S-腺苷甲硫氨酸（AdoMet）依赖性甲基转移酶（MTase）在不同位置自然甲基化。我们报告了负责在起始核苷酸的 N-位置对腺苷进行甲基化的 MTase CAPAM 的共底物范围，该 MTase 利用合成的 AdoMet 类似物进行。化学酶促炔丙基化能够产生特异性修饰的报告 mRNA。这些帽炔丙基化的 mRNA 被高效翻译，并在人细胞中显示出约 3 倍的免疫反应增强。当使用 SARS-CoV-2 的受体结合域（RBD）时，也观察到了相同的效果，SARS-CoV-2 是目前用于 mRNA 疫苗接种的表位之一。因此，真核生物 mRNA 的这种位点特异性化学酶促修饰可能是一种调节 mRNA 翻译和免疫反应的合适策略，可用于未来的治疗应用。
