Fig. 2.

Kinase activity of PknG. The kinase activity of PknG was determined by incubation of 0.1 μg of purified M. bovis BCG and M. smegmatis His-PknG in the absence or presence of the specific PknG inhibitor AX20017 (1 mM) for 30 min at 37°C in a kinase buffer (25 mM Tris/HCl pH 7.4, 2 mM MnCl₂, 1 mM DTT and 3.7 × 10⁵ Bq [γ-³²P]-ATP). The reactions were terminated by the addition of SDS sample buffer and proteins were separated by 10% SDS-PAGE, fixed, dried and autoradiographed to determine the phosphorylation of the PknGs. Results are representative data from one experiment.