Jimbow K, Jimbow M, Chiba M
J Invest Dermatol. 1981 Aug;77(2):213-8. doi: 10.1111/1523-1747.ep12479930.
To characterize how structural properties are related to the morphological differentiation of melanosomes, two forms of melanosomes, commonly seen in mammals, were isolated from B16 and Harding Passey (HP) mouse melanomas. From these morphologically different melanosomes, tyrosinase was solubilized by BRIJ-35 and purified by affinity chromatographies substituted with tyrosinase substrates. We found that tyrosine ethyl ester (TEE) and dopa are effective and specific in retaining tyrosinase as an affinity media and that enzyme retrieval with approximately 100% recovery is possible by a substrate, TEE, or a competitive inhibitor, N-acetyl L-tyrosine. The purified tyrosinase of B16 and HP melanosomes possessed a common antigenic site and revealed little difference in size and the Mikaelis constant for dopa utilization. It is likely that tyrosinase is involved in melanosome morphogenesis only through melanization, and that it is not directly related to the architecture, i.e., lamellar and granular patterns, of the inner matrix.
为了描述结构特性与黑素小体形态分化之间的关系，从B16和哈丁·帕西（HP）小鼠黑色素瘤中分离出两种常见于哺乳动物的黑素小体。从这些形态不同的黑素小体中，用BRIJ - 35溶解酪氨酸酶，并通过用酪氨酸酶底物替代的亲和色谱法进行纯化。我们发现酪氨酸乙酯（TEE）和多巴作为亲和介质在保留酪氨酸酶方面是有效且特异的，并且通过底物TEE或竞争性抑制剂N - 乙酰 - L - 酪氨酸可以实现约100%回收率的酶回收。B16和HP黑素小体的纯化酪氨酸酶具有共同的抗原位点，并且在大小和利用多巴的米氏常数方面几乎没有差异。酪氨酸酶可能仅通过黑素化参与黑素小体的形态发生，并且它与内部基质的结构，即层状和颗粒状模式没有直接关系。
