COMPARISON OF TRPS FROM MURINE AND HUMAN-MALIGNANT MELANOCYTES

Most of our knowledge of the mammalian tyrosinase related protein (TRP ) activities is derived from studies using murine melanoma models, suc h as B16 or Cloudman S-91 melanocytes. Owing to the high degree of hom ology between the murine and human enzymes, it has been assumed that t heir kinetic behavior could be similar. However, the protein sequences at the metal binding sites of the murine and human enzymes show some differences of possible functional relevance. These differences are mo re significant in the metal-A site than in the metal-B site. By using three human melanoma cell lines (HBL, SCL, and BEU), we have studied t he catalytic abilities of the human melanogenic enzymes in comparison to those obtained for the counterpart murine enzymes isolated from B16 melanoma. We have found that TRP2 extracted from all cell lines show dopachrome tautomerase activity although the activity levels in human malignant melanocytes are much lower than in mouse cells. Reconstituti on experiments of the human enzyme indicate that TRP2 has Zn at its me tal binding-sites. Although mouse tyrosinase does not show DHICA oxida se activity, and this step of the melanogenesis pathway is specificall y catalyzed by mouse TRP1, the human enzyme seems to recognize carboxy lated indoles. Thus, human tyrosinase could display some residual DHIC A oxidase activity, and the function of human TRP1 could differ from t hat of the murine protein. Attempts to clarify the nature of the metal cofactor in TRP1 were unsuccessful. The enzyme contains mostly Fe and Cu, but the reconstitution of the enzymatic activity from the apoprot ein with these ions was not possible.