Identification of epitopes on tyrosinase which are recognized by autoantibodies from patients with vitiligo

Abstract

The identification of tyrosinase autoantibodies in some patients with vitiligo has previously been reported. In this study we have determined the B cell epitopes on tyrosinase which are recognized by these autoantibodies. Deletion derivatives of tyrosinase cDNA were constructed and then translated in vitro with the concomitant incorporation of [35S]methionine into the protein products. The 35S-labeled tyrosinase derivatives were subsequently used in radioimmunoassays to investigate the reactivity of sera from five vitiligo patients. The epitope regions identified were: three in a central region of tyrosinase (amino acids 240-255, 289-294, and 295-300) and two others towards the C-terminal end of the protein (amino acids 435-447 and 461-479). Computer analysis of the potential B cell epitopes on tyrosinase revealed that the epitope regions recognized by the vitiligo sera were located in areas predicted to be highly antigenic. In addition, the centrally located antigenic regions (amino acids 289294 and 295-300) had amino acid sequence homology to both tyrosinase-related protein-1 and -2. Thus, the epitopes on tyrosinase recognized by vitiligo patient sera are heterogeneous and include a region with homology to two related proteins which may explain the cross-reactivity previously noted between these antigens.