Regulation of the catalytic activity of preexisting tyrosinase in black and Caucasian human melanocyte cell cultures

Exp Cell Res. 2001 Jan 15;262(2):197-208. doi: 10.1006/excr.2000.5092.

Abstract

The activity of tyrosinase, the rate-limiting enzyme for melanin synthesis, is higher in Black skin melanocytes than in melanocytes derived from Caucasian skin. This variation in enzyme activity is not due to differences in tyrosinase abundance or tyrosinase gene activity, but, rather, is due to differences in the catalytic activity of preexisting tyrosinase. In melanocytes, tyrosinase is localized to the membrane of melanosomes and in Caucasian melanocytes the melanosome-bound enzyme is largely inactive. Conversely, in melanosomes of Black melanocytes, tyrosinase has high catalytic activity. Treatment of Caucasian melanocytes with the lysosomotropic compound ammonium chloride or with the ionophores nigericin and monensin results in a rapid and pronounced increase in tyrosinase activity. This increase occurs without any change in tyrosinase abundance, indicating that these compounds are increasing the catalytic activity of preexisting enzyme. Inhibition of the vacuolar proton pump V-ATPase by treatment of Caucasian melanocytes with bafilomycin also increases tyrosinase activity. In contrast to the 10-fold increase in tyrosinase observed in Caucasian melanocytes, neither ammonium chloride, monensin, nigericin, nor bafilomycin is able to increase the already high level of tyrosinase activity present in melanosomes of melanocytes derived from Black skin. Finally, staining of Caucasian melanocytes with the fluorescent weak base acridine orange shows that melanosomes of Caucasian, but not Black, melanocytes are acidic organelles. These data support a model for racial pigmentation that is based on differences in melanosome pH in Black and Caucasian skin types. The models suggests that melanosomes of Caucasian melanocytes are acidic, while those of Black individuals are more neutral. Since tyrosinase is inactive in an acid environment, the enzyme is largely inactive in Caucasian melanosomes but fully active in Black melanosomes.

MeSH terms

  • Acridine Orange
  • Ammonium Chloride / pharmacology
  • Anti-Bacterial Agents / pharmacology
  • Black People*
  • Blotting, Western
  • Catalysis*
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation / drug effects
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Hydrogen-Ion Concentration / drug effects
  • Macrolides*
  • Male
  • Melanocytes / cytology
  • Melanocytes / drug effects
  • Melanocytes / enzymology*
  • Melanosomes / drug effects
  • Melanosomes / enzymology
  • Monensin / pharmacology
  • Monophenol Monooxygenase / metabolism*
  • Nigericin / pharmacology
  • Skin Pigmentation / physiology
  • White People*

Substances

  • Anti-Bacterial Agents
  • Enzyme Inhibitors
  • Macrolides
  • Ammonium Chloride
  • bafilomycin A
  • Monensin
  • Monophenol Monooxygenase
  • Acridine Orange
  • Nigericin