Effect of Purified Mushroom Tyrosinase on Melanin Content and Melanogenic Protein Expression

Kamal Uddin Zaidi, Sharique A. Ali, Ayesha S Ali
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引用次数: 13

Abstract

In mammalian melanocytes, melanosome is a highly specialized organelle where melanin is synthesized. Melanin synthesis is controlled by tyrosinase, the vital enzyme in melanogenic pathway. The present investigation is based on an effect of purified mushroom tyrosinase of Agaricus bisporus on B16F10 melanocytes for the melanin production via blocking pigment cell machinery. Using B16F10 melanocytes showed that the stimulation of melanogenesis by purified tyrosinase is due to increased tyrosinase absorption. Cellular tyrosinase activity and melanin content in B16F10 melanocytes were increased by purified tyrosinase in a dose-dependent manner. Western blot analysis revealed that cellular tyrosinase levels were enhanced after treatment with purified tyrosinase for 48 hours. Furthermore, tyrosinase induced phosphorylation of cyclic adenosine monophosphate (cAMP) response element-binding protein (CREB) in a dose-dependent manner. The purified tyrosinase-mediated increase of tyrosinase activity was significantly attenuated by H89, LY294002, Ro-32-0432, and PD98059, cAMP-dependent protein kinase inhibitors. The results indicate that purified tyrosinase can be used as contestant for the treatment of vitiligous skin conditions.
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纯化蘑菇酪氨酸酶对黑色素含量和黑色素蛋白表达的影响
在哺乳动物黑素细胞中,黑素小体是一种高度特化的细胞器,用于合成黑色素。黑色素的合成是由酪氨酸酶控制的,酪氨酸酶是黑色素生成途径中的重要酶。本研究是基于纯化双孢蘑菇酪氨酸酶对B16F10黑素细胞的影响,通过阻断色素细胞机制来产生黑色素。使用B16F10黑素细胞表明,纯化酪氨酸酶刺激黑素生成是由于酪氨酸酶吸收增加。纯化的酪氨酸酶使B16F10黑素细胞的酪氨酸酶活性和黑色素含量呈剂量依赖性增加。Western blot分析显示,纯化酪氨酸酶处理48小时后,细胞酪氨酸酶水平提高。此外,酪氨酸酶以剂量依赖的方式诱导环磷酸腺苷(cAMP)反应元件结合蛋白(CREB)的磷酸化。纯化酪氨酸酶介导的酪氨酸酶活性增加被H89、LY294002、Ro-32-0432和PD98059等camp依赖性蛋白激酶抑制剂显著减弱。结果表明,纯化的酪氨酸酶可作为治疗白癜风皮肤病的竞争者。
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