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Figure 1.

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Publication: Stonkus et al., 2025 - A new role for PHYHD1 and related dioxygenases: demethylation of 2'-O-methylated nucleosides
All Figures: Figures for Stonkus et al., 2025

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Figure 1.

Discovery and characterization of FJS catalytic activity. (A) The functionality of FJS, encoded in the pUC19 vector, was identified using the E. coli DH10B ΔpyrFEC strain. (B) The enzymatic activity of FJS depends on 2-oxoglutarate and is further enhanced in vitro by adding ascorbate and FeSO₄. (C) HPLC-MS analysis confirmed the loss of the methyl group during the reaction by detecting a 14 Da decrease in the mass of 2′-O-methylcytidine. (D) The methyl group from the modified nucleoside is converted to formaldehyde, as demonstrated by the reaction with Purpald reagent. (E) Proposed reaction scheme for the FJS-catalyzed demethylation of 2′-O-methylcytidine. Cm, 2′-O-methylcytidine; C, cytidine; 2OG, 2-oxoglutarate; Asc, ascorbate.

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