FIG. 5.

RokA converts NAG to NAG 6-P. (A) Extracts of B. fragilis containing RokA phosphorylate NAG in a time-dependent, ATP-dependent manner (lanes 1 to 4). Extracts were incubated with ATP and [C¹⁴]NAG, as described in Materials and Methods, for the time indicated on each lane. Lanes 5 and 6 were incubated for 60 min at 37°C. Lane 5 is labeled NAG incubated with buffer, ATP, and an extract of the double kinase mutant CJB101. Lane 6 is labeled substrate incubated with CJB200 (ΔhexA) extract and no ATP. Arrows represent standards of NAG and NAG 6-P run on the same chromatogram and developed as described in Materials and Methods.