FIGURE 3.

DHHC1, -3, -7, and -10 (z11) palmitoylate Ncdn. A, 23 individual DHHC proteins were co-expressed with Ncdn, TARP γ-8, CaMKIIα, or Syd-1 in HEK293 cells and metabolically labeled with [³H]palmitate, as shown in Fig. 2A. Arrows indicate the positions of palmitoylated substrates. Palmitoylation of Ncdn was enhanced by DHHC1, -3, -7, and -10 (z11), whereas TARP γ-8, CaMKIIα, and Syd-1 were palmitoylated by the DHHC3/7 subfamily. HA-GST was used for mock indicated as –. B, palmitoylation of Ncdn was increased by the DHHC1/10 (z1/11) and DHHC3/7 subfamilies. The band intensity of palmitoylated Ncdn was quantified and normalized to total Ncdn. The -fold increase in relative band intensity (in the presence of DHHC proteins) to the basal intensity (in the absence of DHHC proteins) is indicated below each lane. The average values of three independent experiments are represented. DHHC1 and -10 (z11) belong to the same subfamily in green, and DHHC3 and -7 belong to the other subfamily in blue. C, expression of 23 HA-DHHC proteins was confirmed by Western blotting (WB) with anti-HA antibody (a representative result).