Figure 5.

(A) Purified CypD (obtained from Abnova) can be S-palmitoylated non-enzymatically in-vitro; unpaired two-tailed Student’s t-test, n = 8 independent replicates per group (each replicate represents an independent reaction) using two different stocks of purified CypD, performed on four different days; representative western blot. S-acylation of CypD was assessed by a resin-assisted capture (Acyl-RAC) that is based on the capture of S-acylated cysteines by a sepharose resin. Inputs represent samples before the Acyl-RAC was run and outputs represent the sample following the Acyl-RAC which is enriched in S-acylated proteins. (B) S-palmitoylation of CypD does not affect its isomerase activity; representative trace and summary of data. CypD isomerase activity was assessed spectrophotometrically; two-way ANOVA with Sidak’s post hoc test, n = 5 independent replicates per group (each replicate represents an independent reaction of in vitro palmitoylation). CsA, cyclosporine A; DMSO blank, spontaneous cis-trans isomerization without CypD; PCoA, palmitoyl-CoA.