Fig. 2.

The Arabidopsis AtPAT10 is auto-acylated. AtPAT10 (PAT10) and AtPAT10C¹⁹²A (PAT10C¹⁹²A) were detected by Western blotting with anti-V5 antibodies by the alkaline phosphatase assay. The lanes in ‘S-acylation’ show the amount of AtPAT10 or AtPAT10C¹⁹²A bound to the neutravidin beads with (+) or without (−) NH₂OH treatment. ‘Loading’ controls showing equal amounts of protein were loaded (10% of the proteins used for ‘S-acylation’ assays). The molecular weight of AtPAT10 and AtPAT10C¹⁹²A is c. 39 kDa. A strong band corresponding to AtPAT10 was detected in the +NH₂OH treated sample in the ‘S-acylation’ lane, indicating that it is bound to an acyl group via a labile thioester linkage confirming that it is auto-acylated. However, no signal was detected for AtPAT10C¹⁹²A and therefore it is not auto-acylated.