Figure 4.
Functional Characterization of MinCN/MinD as Phosphatase/UPRTase
(A) Schematic of MinCN-catalyzed reaction. Udk, uridine kinase (GenBank: QBJ70019.1) from Bacillus subtilis; MIN-5′-MP, MIN-5′-monophosphate.
(B) LC-HRMS analysis of the Udk/MinCN reactions. MIN Std, the authentic standard of MIN; +Udk, the Udk catalyzed reaction to form MIN-MP; -Udk, the reaction without Udk added as negative control; (+Udk)+MinCN, the Udk reaction, proceeded for 4 h, with further addition of MinCN for another 1 h; (+Udk)-MinCN, the Udk reaction (proceeded for 4 h) without further addition of MinCN (also incubated for another 1 h). The characteristic [M + H]+ ions of MIN and MIN-MP are indicated as well in this panel for related peaks.
(C) Schematic of MinD-catalyzed reaction. PRPP, phosphoribosyl pyrophosphate.
(D) HPLC traces of the MinD reactions. Uracil Std, the authentic standard of uracil; UMP Std, the authentic standard of UMP; +MinD, the MinD reaction using uracil and PRPP as substrates; -MinD, the reaction without MinD added as negative control. The characteristic [M + H]+ ions of UMP/uracil are also indicated in this panel.
See also Figures S6 and S7; Tables S1 and S6.