A benzylic linker promotes methyltransferase catalyzed norbornene transfer for rapid bioorthogonal tetrazine ligation

Abstract

Benzylic AdoMet analogs enable highly efficient enzymatic transfer of norbornenes to nucleic acids and subsequent tetrazine ligation.

Efficient cosubstrate enzyme pairs for sequence-specific methyltransferase-directed photolabile caging of DNA

“…A cause for this effect may be the enhanced reactivity of benzylic positions towards substitution and the conjugative stabilization of the sp 2 -hybridized S N 2-transition state through the aromatic system, which has recently also been observed for RNA MTases. 28 Irradiating samples of caged duplex ODNI R ÁODNII Me for different time periods at l = 365 nm using a UV hand lamp (6 W) revealed clean and rapid photorelease of uncaged duplex for NB-, 4MNB-and 5MNB-modified ODNI and thus confirmed the reversibility of the enzymatic labelling reaction (Fig. 2d and Fig.…”

Covalent labeling of nucleic acids

“…11), as they enabled a fast tetrazine ligation for post-synthetic labeling of DNA and RNA. 147 Lately, AdoMet analogs were also used to modify DNA with photocleavable groups. 148 Two main strategies have been established to label DNA using MTases and fluorophore bearing cofactors.…”

Synthetic mRNA capping

“…Recently, we revealed that the N 7-cap methyltransferase Ecm1 from Encephalitozoon cuniculi is highly promiscuous. Sterically very demanding AdoMet analogues bearing for example a norbornene or 4-vinylbenzyl moiety were efficiently converted [ 92 – 93 ]. The pronounced promiscuity can be attributed to the structure of Ecm1 which forms a substrate binding cleft rather than a pocket [ 94 ].…”