Muttach F, Muthmann N, Reichert D, Anhäuser L, Rentmeister A
University of Münster , Department of Chemistry , Institute of Biochemistry , Wilhelm-Klemm-Str. 2 , 48149 Münster , Germany . Email:
Cells-in-Motion Cluster of Excellence (EXC1003-CiM) , University of Münster , Germany . Email:
Chem Sci. 2017 Dec 1;8(12):7947-7953. doi: 10.1039/c7sc03631k. Epub 2017 Oct 10.
Site-specific alkylation of complex biomolecules is critical for late-stage product diversification as well as post-synthetic labeling and manipulation of proteins and nucleic acids. Promiscuous methyltransferases in combination with analogs of -adenosyl-l-methionine (AdoMet) can functionalize all major classes of biomolecules. We show that benzylic moieties are transferred by Ecm1 with higher catalytic efficiency than the natural AdoMet. A relative specificity of up to 80% is achieved when a norbornene moiety is placed in -position, enabling for the first time enzymatic norbornene transfer to specific positions in DNA and RNA- even in cell lysate. Subsequent tetrazine ligation of the stable norbornene moiety is fast, efficient, biocompatible and - in combination with an appropriate tetrazine - fluorogenic.
复杂生物分子的位点特异性烷基化对于后期产物多样化以及蛋白质和核酸的合成后标记与操作至关重要。混杂型甲基转移酶与S-腺苷-L-甲硫氨酸（AdoMet）类似物相结合，可使所有主要类别的生物分子功能化。我们发现，Ecm1转移苄基部分的催化效率高于天然AdoMet。当降冰片烯部分位于α-位时，可实现高达80%的相对特异性，这首次使得酶促降冰片烯转移至DNA和RNA中的特定位置——即使是在细胞裂解物中。稳定的降冰片烯部分随后的四嗪连接反应快速、高效、具有生物相容性，并且与合适的四嗪结合时会产生荧光。
