Tyrosinase, the key enzyme in melanogenesis, is activated when protein kinase C-beta (PKC-beta) phosphorylates the serine residues at amino acid positions 505 and 509. To further elucidate the mechanism by which phosphorylation of tyrosinase by PKC-beta leads to the activation of tyrosinase, a possible complex formation between phosphorylated tyrosinase and tyrosinase related protein-1 (TRP-1), a melanogenic protein suggested to influence tyrosinase activity, was investigated. Non-denaturing gel electrophoresis of melanocyte lysate revealed two molecular weight forms of TRP-1 and a monoclonal antibody against TRP-1 co-immunoprecipitated tyrosinase and TRP-1, suggesting that TRP-1 may be complexed with tyrosinase. Activation of PKC by treating melanocytes with phorbol 12,13-dibutyrate (PDBu) increased the level of tyrosinase co-immunoprecipitated with TRP-1; whereas a selective PKC inhibitor bisindolylmaleimide inhibited PDBu-induced increase in the level of tyrosinase co-immunoprecipitated with TRP-1. These results suggest that phosphorylation of tyrosinase by PKC-beta induces a complex formation between tyrosinase and TRP-1.