Synthesis and degradation of tyrosinase in cultured melanoma cells.
The tyrosinase (EC 1.14.18.1) activity of cultured B-16 mouse melanoma cells (C2M) in the stationary phase depends greatly on whether the culture medium contains glucose or galactose. The activity in medium containing galactose was about ten times that in medium containing glucose at pH 7.2. This difference in tyrosinase activity was concluded to be due to a shift of balance between synthesis and degradation of the enzyme. Experiments were conducted with stationary phase cultures in the presence of cycloheximide. The melanoma cells did not synthesize tyrosinase in medium containing glucose in the stationary phase. But when they were cultured under identical conditions, except that glucose was replaced by galactose, they continued to synthesize tyrosinase. The rate of synthesis in medium containing galactose at pH 6.3 was one third of that in the same medium at about pH 7, in which the increase in specific activity of tyrosinase per day was about 30 nmoles/mg cell protein per hr. The rate of degradation of the enzyme was practically the same in medium containing glucose as in medium containing galactose, and largely depended on the pH of the culture medium. At pH 6.3, the half-life was about one third of that at pH 7.2, where it was about 1.8 days. The degradation at acidic pH values was much reduced by ammonium salt and was strongly inhibited by the protease inhibitor, leupeptin.
D000643:Ammonium Chloride / Q000494:pharmacology; D000818:Animals; D004156:Catechol Oxidase / Q000378:metabolism*; D002460:Cell Line; D003470:Culture Media; D005690:Galactose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007976:Leupeptins / Q000494:pharmacology; D008545:Melanoma; D051379:Mice; D014442:Monophenol Monooxygenase / Q000378:metabolism*
D003470:Culture Media; D007976:Leupeptins; D000643:Ammonium Chloride; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase; D005947:Glucose; D005690:Galactose