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. 2019 Nov 14;118(4):980–988. doi: 10.1016/j.bpj.2019.11.003

Figure 2.

Figure 2

Functional reconstitution of hDHHC20 in a POPC bilayer. (A) An experimental assay of the autoacylation activity of hDHHC20 in POPC nanodiscs, either in the presence of acyl-CoA (closed circles) or in its absence (open circles), is shown. Autoacylation of hDHHC20 is spontaneous; addition of DTT breaks the thioester linkage between Cys156 and acyl-CoA, thereby regenerating the enzyme from the autoacylated form and enabling multiple rounds of catalysis. (B) Impairment of the autoacylation reaction assayed in (A) by addition of 2-BP, a competitive inhibitor of hDHHC20, is shown. (C) An all-atom molecular dynamics simulation system, including hDHHC20 (yellow surface), a POPC bilayer (gray/blue/orange/red lines), water (blue lines), and 100 mM NaCl (green/magenta spheres), is shown. The figures show the final snapshot of one of the two calculated trajectories, each 1 μs long. To see this figure in color, go online.