Figure 3. In vitro activity of MurT and GatD.

(A) Purified recombinant GatD and MurT enzymes were incubated with purified lipid II. In the negative control (lane 1) the reaction was immediately stopped by the addition of BuOH/Pyr/Ac. Time dependency of the GatD/MurT catalyzed reaction. (B) Purified [¹⁴C]-lipid II was incubated in the presence of 2 µg MurT and GatD each, glutamine and ATP. The reaction was stopped by the addition of BuOH/PyrAc and the amount of amidated lipid II was quantified using phosphoimaging; mean values of three independent experiments are given.