Regulation of mammalian melanogenesis. I: Partial purification and characterization of a dopachrome converting factor: dopachrome tautomerase.
A protein that catalyzes the decoloration of dopachrome has been partially purified from B16 mouse melanoma tumors. The enzyme is preferentially associated to the melanosomes, but it is also found in the microsomal and cytosolic fractions of cellular homogenates. The protein is clearly different from tyrosinase, and should be related to the dopachrome oxidoreductase (Barber et al. (1984) J. Invest. Dermatol. 83, 145-149) and the dopachrome conversion factor (Korner and Pawelek (1980) J. Invest. Dermatol. 75, 192-195) since the reaction product of dopachrome conversion is 5,6-dihydroxyindole-2-carboxylic acid. The protein appears to have an oligomeric structure, with a molecular mass slightly higher than 300 kDa estimated by gel filtration, whereas the molecular mass of the monomer might be approx. 46 kDa estimated by SDS-PAGE electrophoresis. Its Km for dopachrome is around 100 microM. The enzyme is competitively inhibited by indoles and is unaffected by metal chelators. It also has the ability to increase the amount of melanin formed from L-tyrosine by melanoma tyrosinase, and therefore, cannot be considered an 'indole blocking factor' as was suggested for the related dopachrome oxidoreductase. Since the reaction catalyzed by the enzyme is a tautomeric shift on dopachrome, we would propose dopachrome tautomerase (EC 5.3.2.3) as the most precise and informative name.
Aroca
P
P
Departamento de Bioquímica y Biología Molecular, Facultad de Medicina, Universidad de Murcia, Spain.
Garcia-Borron
J C
JC
Solano
F
F
Lozano
J A
JA
D000818:Animals; D004156:Catechol Oxidase / Q000302:isolation & purification* / Q000378:metabolism; D002460:Cell Line; D002845:Chromatography; D002850:Chromatography, Gel; D017886:Durapatite; D005260:Female; D006882:Hydroxyapatites; D019746:Intramolecular Oxidoreductases; D007700:Kinetics; D008297:Male; D008543:Melanins / Q000096:biosynthesis*; D008546:Melanoma, Experimental / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008970:Molecular Weight; D013347:Subcellular Fractions / Q000201:enzymology
D006882:Hydroxyapatites; D008543:Melanins; D017886:Durapatite; D004156:Catechol Oxidase; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase