Figure 3. MEND is blocked by preventing protein palmitoylation reactions.
(A) From left to right, composite results for Standard MEND after incubating cells with 1:1 palmitate-loaded albumin (50 μM) for 1 hr (black), after incubating cells with fatty acid-free albumin (Alb, 50 μM) for 1 hr (purple), with the palmitoylation inhibitor, bromopalmitate (BP, 50 μM) included in all solutions (blue), with the acyl CoA synthetase inhibitor, Triascin C (TrC, 2 μM) in all solutions (green), with BP (50 μM) and TrC (2 μM) in all solutions (orange), with cytoplasmic acetate (Ace, 6 mM; pink), with acetate (6 mM) and acetyl CoA synthetase (ACS, 20 μM; yellow), and with a high cytoplasmic CoA concentration (3 mM) to block DHHCs (red). For all results, n > 6. (B) CoA inhibition of DHHC2-mediated palmitoylation of the N-terminus of myristoylated lymphocyte-specific kinase (myrLckNT). For details, see ‘Materials and methods’.