Fig. 4.

Reaction of Cpr19 and LipL with the phosphonate derivative of UMP. (A) Proposed outcome for the reaction of 5′-deoxyuridine-5′-methylphosphonate (UMcP) with LipL and Cpr19. Amino acid ligands denote protein side chains used to bind the iron and co-product succinate. (B) Lineweaver-Burk plot of UMcP inhibition with respect to variable UMP with LipL. (C) HPLC analysis of the Cpr19-catalyzed reaction starting with UMcP in comparison to synthetic standards (5′S)-hydroxy-UMcP and (5′R)-hydroxy-UMcP and control reactions without enzyme. *denotes an unidentified contaminant of synthetic 5′-deoxyuridine-5′-methylphosphonate with a distinct UV-VIS profile compared to uracil-containing compounds; U denotes uracil. A₂₆₀, absorbance at 260 nm.