Figure 7.
Evidence for formation of the TreS:Pep2 complex in M. smegmatis mc2155. (A) His6-tagged Pep2 was purified from M. smegmatis extracts by Ni-NTA and ion exchange chromatography, and anion exchange column fractions were analyzed by Coomassie-stained SDS-PAGE, with NaCl concentrations (in M, across the top), and molecular weight standards (in kDa, on the left) as indicated. FT = flow through. (A, inset) Coomassie-stained SDS-PAGE of ion exchange fractions eluted at 0.35 and 0.4 M NaCl after ∼20-fold concentration, flanked by TreS- and Pep2 controls. (B) Thin-layer chromatogram of reactions mixtures (100 μL) containing 100 mM trehalose (Tre) and 20 μL of fractions of the anion exchange column fractions shown in panel A. The position of maltose-1-phosphate (M1P) and NaCl concentrations (in M) are indicated. Arrows highlight the two fractions producing maltose-1-phosphate.