TET‐Like Oxidation in 5‐Methylcytosine and Derivatives: A Computational and Experimental Study

The epigenetic marker 5‐methylcytosine (5mC) is an important factor in DNA modification and epigenetics. It can be modified through a three‐step oxidation performed by ten‐eleven‐translocation (TET) enzymes and we have previously reported that the iron(IV)‐oxo complex [Fe(O)(Py(5)Me(2)H)](2+) (1) can oxidize 5mC. Here, we report the reactivity of this iron(IV)‐oxo complex towards a wider scope of methylated cytosine and uracil derivatives relevant for synthetic DNA applications, such as 1‐methylcytosine (1mC), 5‐methyl‐iso‐cytosine (5miC) and thymine (T/5mU). The observed kinetic parameters are corroborated by calculation of the C−H bond energies at the reactive sites which was found to be an efficient tool for reaction rate prediction of 1 towards methylated DNA bases. We identified oxidation products of methylated cytosine derivatives using HPLC‐MS and GC‐MS. Thereby, we shed light on the impact of the methyl group position and resulting C−H bond dissociation energies on reactivity towards TET‐like oxidation.