Unravelling 2-oxoglutarate turnover and substrate oxidation dynamics in 5-methylcytosine-oxidising TET enzymes.

ABSTRACT: Fe(II)- and 2-oxoglutarate (2OG)-dependent dioxygenases use 2OG and O₂ cofactors to catalyse substrate oxidation and yield oxidised product, succinate, and CO₂. Simultaneous detection of substrate and cofactors is difficult, contributing to a poor understanding of the dynamics between substrate oxidation and 2OG decarboxylation activities. Here, we profile 5-methylcytosine (^5mC)-oxidising Ten-Eleven Translocation (TET) enzymes using MS and ¹H NMR spectroscopy methods and reveal a high degree of substrate oxidation-independent 2OG turnover under a range of conditions. 2OG decarboxylase activity is substantial (>20% 2OG turned over after 1 h) in the absence of substrate, while, under substrate-saturating conditions, half of total 2OG consumption is

SUBMITTER: Simelis K 

PROVIDER: S-EPMC11662004 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature