Analysis of tyrosinases as asparagin-linked oligosaccharides by concanavalin A lectin chromatography: appearance of new segment of tyrosinases in melanoma cells following interrupted melanogenesis induced by glycosylation inhibitors.
The structural alteration of carbohydrate moieties of tyrosinases associated with the depigmentation process induced by glycosylation inhibition has been investigated by using concanavalin A (Con A) affinity chromatography. Con A affinity chromatography of deoxycholate-solubilized large and small granule fractions shows that while all tyrosinases found in control B-16 cells exhibit affinity to Con A lectin, there is an emergence on non-Con A binding tyrosinases in the unpigmented cells induced by glycosylation inhibitors, such as tunicamycin and glucosamine. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, control tyrosinase activity forms 2 distinct bands consisting of T1 and T3. But tyrosinases from the unpigmented cells lose T3 tyrosinase and are resolved into a few different molecular weight components, one of which is Con A affinitive T1 tyrosinase and the others are non-Con A affinitive tyrosinases with smaller molecular weights than the T1 tyrosinase. These findings suggest that altered structures of carbohydrate moiety in tyrosinase molecules play a role in the induction of loss of membrane-binding capacity of tyrosinases, resulting in the loss of melanization in pigment cells.
D000818:Animals; D002241:Carbohydrates / Q000096:biosynthesis; D004156:Catechol Oxidase / Q000032:analysis*; D002846:Chromatography, Affinity; D003208:Concanavalin A / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D008543:Melanins / Q000096:biosynthesis; D008545:Melanoma / Q000032:analysis*; D051379:Mice; D014442:Monophenol Monooxygenase / Q000032:analysis*; D014415:Tunicamycin / Q000494:pharmacology
D002241:Carbohydrates; D008543:Melanins; D003208:Concanavalin A; D014415:Tunicamycin; D003513:Cycloheximide; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase