Figure 2.
ShHTL7 interacts with AtMAX2 and AtSMAX1 with high SL sensitivity and low SL specificity. (A) The structures of the four natural SLs used in this and subsequent assays. STR and ORO were used as racemic mixtures. (B) The interactions between ShHTLs and AtMAX2 or AtSMAX1 separately in the presence of 1 µM of rac-GR24, 5DS, 4DO, STR, and ORO were assessed in Y2H assays. AtMAX2 and AtSMAX1 were used as prey fused with Gal4 AD AD-AtMAX2 and AD-SMAX1 and ShHTL proteins were fused with the BD (BD-ShHTL). DMSO was used as solvent for SLs and as negative control in assays. (C) The Y2H assay was used to detect the binding of selected BD-ShHTLs with AD-AtMAX2 in the presence of 10−7 to 10−10 M rac-GR24, 5DS, 4DO, STR, and ORO. Interactions were detected in the control medium and the selective medium SD/-Leu/-Trp/-His/-Ade (-His, Ade).