Purification and Characterization of Glutathione Binding Protein GsiB from Escherichia coli

Figure 2

Native gel and SDS-PAGE analysis of GsiB binding activity with GSH and GSSG. (a) Native gel analysis of GsiB protein with GSH and GSSG. M: Marker; Lane 1: purified GsiB protein; Lane 2: GsiB incubated with GSH; Lane 3: GsiB incubated with GSSG. (b) SDS-PAGE analysis of GsiB protein with GSH and GSSG. M: Marker; Lanes 1-4: GsiB protein incubated at 25°C for 2 h; Lane 5-7: same protein aliquots as Lanes 1-4 incubated with GSH (Lanes 5-6) and GSSG (Lane 7-8). Lanes 1, 3, 5, and 7 were GsiB stored in -80°C for 6 months. Lanes 2, 4, 6, and 8 were freshly purified GsiB. (c) Western blot analysis of purified GsiB. Lane 1: purified GsiB protein; Lane 2: GsiB protein with GSH.

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