Extended Data Fig. 7: PolE activity assays.
From: Azetidine amino acid biosynthesis by non-haem iron-dependent enzymes
(a) LC-MS analysis (EIC at m/z 363.1373) of PolE assays. PolE assays were performed with 150 μM l-Ile, 15 μM PolE, 100 μM Fe(II), 1 mM BH4, 1 mM ascorbate and ~0.5 mM O2 (i). Also shown are controls without Fe (ii), without BH4 (iii), without O2 (iv), and without PolE (v). (b) LC-MS analysis of PolE assays with different cofactors (EIC at m/z 363.1373 and 365.1530). The assays were performed with 150 μM l-Ile, 15 μM PolE, 100 μM Fe(II), 1 mM ascorbate, and ~0.5 mM O2, with 1 mM α-KG (i), with 1 mM THF (ii), with 1 mM BH4 (iii), with 100 μM FAD and 1 mM NADH mimic (iv), and with 100 μM FMN and 1 mM NADH mimic (v). Source data are provided.
