VieB does not disrupt the GST-VieS-C/VieA-His
interaction. GST-VieS-C (lane 5–10) or GST alone (lane 11–14) bound to glutathione beads were incubated with either, pre-mixed 5 μM VieA-His6 and 25 μM VieB (VieB), 5 μM VieA-His6 alone (VieA) or 5 μM VieA-His6 and 25 μM BSA (BSA). Reactions were washed with 150 μl of wash buffer five times and protein was eluted off the beads with 150 μl of elution buffer containing reduced glutathione. Samples were taken from the input (I), wash (W) and elution (E) fractions for analysis on 10% SDS Page gel stained with Lumitein protein stainTM. VieB, VieA and BSA inputs are indicated by lanes 2–4, respectively. Percentage of VieA-His6 bound is the average of four replicates with standard deviation shown below; gel shown is a representative of the replicates. Statistical significance was determined by one-way ANOVA and Dunn’s Multiple Comparison. M = protein standard.