Fig. 2

The N-terminal region of HCN2 is the site for S-palmitoylation. a Schematic structure of the HCN subunit and location of intracellular cysteines in the HCN2 subtype. Each HCN channel subunit contains six transmembrane segments (S1–S6) and a C-terminal 120-aa cyclic nucleotide binding domain (CNBD). There are ten intracellular cysteines in HCN2, of which only three (C508, C584, and C601) located in the C-linker and CNBD are conserved in the HCN family channels. b Effects of various cysteine site mutations on S-palmitoylation of HCN2 (see text for a description of the mutants). Representative blot shows the abolishment of S-palmitoylation in the ΔN125 mutant, in which all five cysteine sites in the N-terminus of HCN2 were eliminated. Normalized intensities were calculated as in Fig. 1, and are shown below (n = 3). ***p < 0.001; ns not significant versus HCN2