Fig. 3

All five N-terminal cysteines may equally contribute to the S-palmitoylation of HCN2. Representative blots showing the results of the ABE assay for HCN2 proteins containing various N-terminal cysteine-site mutations (see text for a description of the mutants). Although not statistically significant, we observed a 10–20 % decrease in biotin-labeling for the N-terminal single cysteine-site mutants (C63A and C69A in A, and C82A, C89A, and C104A in b). By contrast, the biotin signal of the ΔCysNT mutant protein (all five N-terminal cysteines were replaced with alanines) was at undetectable levels. Normalized intensities were calculated as in Fig. 1, and are shown below (n = 3). ***p < 0.001; ns not significant versus HCN2