Figure 6 | Scientific Reports

Figure 6

From: Functional characterization and discovery of modulators of SbMATE, the agronomically important aluminium tolerance transporter from Sorghum bicolor

Figure 6

Purified SbMATE binds ethidium bromide, but not propidium iodide. (A) Detergent purified SbMATE elutes from size exclusion chromatography as a symmetrical monodispersed peak (FPLC gel filtration trace on left) that is deemed to be ~95% homogeneous (coomassie stained SDS-PAGE of pooled FPLC peak fractions shown on right). (B) Purified SbMATE was covalently immobilized on a CMD surface plasmon resonance (SPR) sensorchip. 100 µL of varying concentrations of ethidium bromide were injected, with each concentration having an association and dissociation time of 120 s. The chip was regenerated between each injection using a short 10 µL injection of 50 mM glycine-HCl, pH 2.1. Traces on the sensorgram represent double-referenced data, obtained by subtracting response signals from a blank surface and buffer injections (n = 5 replicates performed using independently purified batches of protein, traces represent data from a single experiment). (C) SPR was conducted as described in Fig. 6B, except with varying concentrations of propidium iodide instead of ethidium bromide (n = 3 independent replicates, sensorgrams are from a single experiment).

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