Figure 2
From: The dual roles of RPE65 S-palmitoylation in membrane association and visual cycle function
Mutational analysis of cysteine residues to identify potential palmitoylation sites in RPE65. (A) Schematic of dog RPE65 amino acid sequence showing cysteine residues (numbers in superscript represent the residue position). Cysteines in red and blue colour indicate the non-conserved and conserved cysteines, respectively, among different species. (B) Identification of palmitoylated cysteine residues. HEK293F cells overexpressing mutant C112A, C146A and C195A proteins were analysed for palmitoylation using acyl-RAC assay followed by immunoblotting for RPE65 using ~20 µg of input and pulldown eluted samples. Samples were treated with 0.5 M hydroxylamine (HAM; indicated as “+”) or 0.5 M NaCl (indicated as “−”), respectively. Results were calculated as mean ± S.D. from three independent experiments. *P < 0.0001, **P < 0.0003, unpaired student’s t-test.
