Figure 6.

Activity of TreS and Pep2 and evidence for TreS affecting Pep2 activity. (A) Thin layer chromatography analysis of reaction products demonstrating Pep2-catalyzed conversion of maltose to maltose-1-phosphate and conversion of trehalose to maltose-1-phosphate when both TreS and Pep2 are present. (B,C) Michaelis–Menten kinetics of M. tuberculosis Pep2 activity, monitored by an enzyme-coupled assay (see Methods). Experiments were done in triplicate and Michaelis–Menten parameters (Table 2) determined using the GraphPad Prism software. (D) Hydrolysis activity of M. smegmatis TreS (0.4 μM) in the absence and presence of M. smegmatis Pep2-K145A. Molar ratios of Pep2-K145A:TreS are indicated, and data points for [TreS] = 0.8 μM (Figure S6C, Supporting Information) are included for comparison. Data points were fitted to a linear function (v_i = k[Tre]).