pH-dependent interconversion of two forms of tyrosinase in human skin.
1. We have shown that the characteristic lag in cresolase activity of human skin tyrosinase at inhibitory concentration of tyrosine was absent at all pH values studied, i.e. pH 5.2, 5.7, 6.2 and 6.8, if the enzyme solubilized at low pH was used as the source of enzyme, but the same enzyme when dialysed against buffers of various pH values showed linear activity only at pH 5.2 and was not inhibited by excess tyrosine, whereas at higher pH values it exhibited a lag and inhibition by excess tyrosine. 2. However, the enzyme solubilized in buffer/detergent, pH 6.8, when dialysed against buffer of the same pH showed linear activity at pH 5.2 and non-linear activity at pH 6.8. 3. The water/detergent-solubilized enzyme from human skin melanosomes showed linear activity even at inhibitory concentrations of tyrosine at pH 5.2 and 6.8 up to 2 h, but acceleration of rate was observed after 2 h for the enzyme measured at pH 6.8. 4. After dialysis of the water/detergent-solubilized enzyme against double-glass-distilled water, it still exhibits linear activity at inhibitory concentration of tyrosines at pH 6.8 for the first 2 h, but the same enzyme when dialysed against 0.02 M-sodium phosphate buffer, pH 6.8, exhibits negligible activity up to 1/2 h, in contrast with considerable activity before dialysis during the same interval of time, but without any loss of activity at later intervals of incubation time. 5. On the basis of these results, it is concluded that the enzyme exists in at least two interconvertible forms, one without lag and inhibition by excess tyrosine and the other with lag and inhibition by excess tyrosine. These two forms are interconvertible only by gradual change in pH over a period of hours.
D004156:Catechol Oxidase / Q000378:metabolism*; D002951:Citrates / Q000494:pharmacology; D019343:Citric Acid; D003902:Detergents / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism*; D008544:Melanocytes / Q000187:drug effects / Q000201:enzymology; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D012867:Skin / Q000201:enzymology*; D014443:Tyrosine / Q000378:metabolism
D002951:Citrates; D003902:Detergents; D007527:Isoenzymes; D019343:Citric Acid; D014443:Tyrosine; D004156:Catechol Oxidase; D014442:Monophenol Monooxygenase