Fig. 2.

Rapid turnover of Lck palmitate in unstimulated cells. (A) Schematic of 17-ODA metabolic labeling and detection of palmitoylated proteins using the click chemistry reaction. (B) Lck palmitate turnover kinetics. Jurkat cells were incubated with 1 μM 17-ODA for the indicated times. Palm-Lck, palmitoylated Lck detected by infrared imaging of labeled 17-ODA; IB, immunoblot of total Lck. (C) Fyn palmitate turnover kinetics. (Left) No labeling was noted after 15 min of 17-ODA incubation at room temperature. (Right) Labeling became evident after 1 h of incubation at 37 °C. (D) Lck palmitate turnover kinetics in the presence of APT1 inhibitor palmostatin B. Jurkat cells were pretreated with 10 μM palmostatin B or DMSO for 30 min before the addition of 1 μM 17-ODA.