Palumbo A, d'Ischia M, Misuraca G, Prota G
Stazione Zoologica University of Naples, Italy.
Biochim Biophys Acta. 1991 Jan 23;1073(1):85-90. doi: 10.1016/0304-4165(91)90186-k.
Hydroquinone (HQ) is one of the most effective inhibitors of melanogenesis in vitro and in vivo, and is widely used for the treatment of melanosis and other hyperpigmentary disorders. In an attempt to get some insight into the molecular mechanism of the depigmenting action, which is still very poorly understood, we have investigated the effect of HQ on the tyrosinase catalysed conversion of tyrosine to melanin. Incubation of 0.5 mM tyrosine with 0.07 U/ml tyrosinase in phosphate buffer at pH 6.8 in the presence of 0.5 mM HQ led to no detectable melanin formation, due to the preferential oxidation of HQ with respect to tyrosine (HPLC evidence). Kinetic investigations showed that HQ is a poorer substrate of tyrosinase than tyrosine; yet, it may be effectively oxidised in the presence of tyrosine owing to the generation of catalytic amounts of dopa acting as cofactor of tyrosinase. Product analysis of HQ oxidation with tyrosinase in the presence of dopa showed the predominant formation in the early stages of hydroxybenzoquinone (HBQ), arising from enzymic hydroxylation and subsequent oxidation of HQ, along with lower amounts of benzoquinone (BQ). These results suggest that the depigmenting activity of HQ may partly be related to the ability of the compound to act as an alternate substrate of tyrosinase, thereby competing for tyrosine oxidation in active melanocytes.
对苯二酚(HQ)是体外和体内最有效的黑素生成抑制剂之一,广泛用于治疗黑变病和其他色素沉着过多症。为了深入了解其脱色作用的分子机制(目前对此仍知之甚少),我们研究了HQ对酪氨酸酶催化酪氨酸转化为黑色素的影响。在pH 6.8的磷酸盐缓冲液中,0.5 mM酪氨酸与0.07 U/ml酪氨酸酶在0.5 mM HQ存在下孵育,未检测到黑色素形成,这是由于HQ相对于酪氨酸优先被氧化(HPLC证据)。动力学研究表明,HQ作为酪氨酸酶的底物比酪氨酸差;然而,由于生成了作为酪氨酸酶辅因子的催化量多巴,它在酪氨酸存在下可能被有效氧化。在多巴存在下,酪氨酸酶氧化HQ的产物分析表明,在早期主要形成羟基苯醌(HBQ),它由HQ的酶促羟基化和随后的氧化产生,同时还有少量的苯醌(BQ)。这些结果表明,HQ的脱色活性可能部分与该化合物作为酪氨酸酶替代底物的能力有关,从而在活跃的黑素细胞中竞争酪氨酸氧化。