FIGURE 3.

Protein acetylation in mycobacteria and identification of USP as an interacting partner of MSMEG_5458. A, whole cell lysates (50 μg) prepared from cultures of M. tuberculosis H37Rv, M. bovis Bacillus Calmette-Guérin, and M. smegmatis were separated by SDS-gel electrophoresis and Western blotting performed with acetyl-lysine antibodies in the presence or absence of acetylated bovine serum albumin. B, GST or GST-MSMEG_5458 bound to glutathione beads were incubated with cytosolic fractions prepared from M. smegmatis, in the presence or absence of 10 μm cAMP. Interacting proteins were resolved by SDS-PAGE and visualized by staining with Coomassie Brilliant Blue. One protein (*) was analyzed by mass spectrometry and identified to be MSMEG_4207 (USP). C, samples obtained after pulldown experiments under the conditions indicated were analyzed by Western blotting with acetyl-lysine antibodies. Experiments were repeated thrice.