Figure 5

Arabidopsis AGG2 is S-acylated. Total membranes were purified from Arabidopsis plants expressing a FLAG-YFP-AGG2 fusion from the 35S promoter and subjected to the biotin switch assay. Samples were treated with (Hyd+) or without (Hyd-) the thioester cleavage reagent hydroxylamine. The loading controls show that equal amounts of AGG2 were loaded onto neutravidin beads. The lanes labelled 'S-acylation' show the levels of AGG2 recovered from neutravidin beads, and therefore originally S-acylated. FLAG-YFP-AGG2 has a mass of approximately 40 kDa. The results indicate that AGG2 is S-acylated in planta.