Figure 3 | Scientific Reports

Figure 3

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

Figure 3

Substitution D127N impairs ethidium transport by SbMATE. (A) Sequence alignment of SbMATE with well characterized bacterial NorM from V. Cholera (top row) and plant MATEs, highlighting conservation of a catalytically important aspartate residue (indicated by the red arrow) across bacteria and plant MATEs associated with organic acid transport. (B) Western blot (left) and coomassie stained SDS-PAGE (right) analysis of equal amounts of prepared Pichia pastoris plasma membranes expressing equivalent amounts of either SbMATE D127N, SbMATE WT, or alternatively transformed with empty vector (C) Ethidium transport was conducted as described in Fig. 2A except SbMATE WT was replaced by SbMATE WT-3C-His (see main text for details). SbMATE WT-3C-His transports ethidium efficiently (n = 3 independent replicates, mean ± SEM). (D) Ethidium transport was performed as per Fig. 2A, except with induced cells transformed with SbMATE WT-3C-His, SbMATE D127N, or vector control. Transport rates were calculated as described in Fig. 2C, with the rate for SbMATE WT-3C-His set to 100% (n = 3 independent replicates, data are mean ± SEM).

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