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LC–MS analysis of 2-HPP-CMP from SUMO-Fom3 reactions with CD3-SAM and/or CD3Cbl under (a) multiple-turnover conditions (20 μM SUMO-Fom3 and 1 mM 2-HEP-CMP) and (b) single-turnover conditions (100 μM SUMO-Fom3 and 50 μM 2-HEP-CMP). Formation of the unlabeled product in single-turnover reactions with CD3-SAM and no added MeCbl demonstrates that as-isolated SUMO-Fom3 contains transferable methyl groups.
