Carstam R, Hansson C, Rorsman H, Rosengren E
Acta Derm Venereol. 1986;66(5):369-74.
The dopa oxidase activity of tyrosinase in the skin from albino and black mice was assayed using a technique based on the formation of two diastereomers of 5-S-cysteinyldopa when incubating tissue extracts with both L-dopa and D-dopa as substrates in the presence of cysteine. The amounts of 5-S-L-cysteinyl-L-dopa and 5-S-L-cysteinyl-D-dopa formed were then determined. In the extract from black mouse skin the L-L-diastereomer was produced in more than ten times the amount of the L-D-diastereomer. This stereospecific dopa-oxidation is indicative of the presence of tyrosinase and corresponds well with earlier determinations of the rates of oxidation for human tyrosinase, using L-dopa and D-dopa as substrates. Stereospecific dopa oxidation was absent in albino skin, and the nonspecific dopa-oxidation was two orders of magnitude less than the dopa oxidation in black skin. The study demonstrates the lack of tyrosinase activity in albino skin, and quantifies the non-specific dopa oxidation. The lack of tyrosinase activity in the eluates from albinotic skin was found not to be due to the presence of a tyrosinase inhibitor.
采用一种技术测定白化小鼠和黑色小鼠皮肤中酪氨酸酶的多巴氧化酶活性,该技术基于在半胱氨酸存在下,将组织提取物与L-多巴和D-多巴作为底物一起孵育时形成5-S-半胱氨酰多巴的两种非对映异构体。然后测定形成的5-S-L-半胱氨酰-L-多巴和5-S-L-半胱氨酰-D-多巴的量。在黑色小鼠皮肤提取物中,L-L-非对映异构体的生成量是L-D-非对映异构体的十多倍。这种立体特异性的多巴氧化表明存在酪氨酸酶,并且与早期以L-多巴和D-多巴为底物测定人酪氨酸酶氧化速率的结果非常吻合。白化小鼠皮肤中不存在立体特异性多巴氧化,且非特异性多巴氧化比黑色皮肤中的多巴氧化低两个数量级。该研究证明了白化小鼠皮肤中缺乏酪氨酸酶活性,并对非特异性多巴氧化进行了定量。发现白化小鼠皮肤洗脱液中缺乏酪氨酸酶活性并非由于存在酪氨酸酶抑制剂。