Quevedo W C, Holstein T J, Bienieki T C
Proc Soc Exp Biol Med. 1975 Dec;150(3):735-40. doi: 10.3181/00379727-150-39116.
The T1 variety of tyrosinase is present in both particulate and soluble or readily solubilized forms in the pigmented hypodermis (hair bulbs) of C57BL mice and Harding-Passey mouse melanoma. Trypsin treatment of 35,000g supernatants containing the microsomal (small granule) fraction of gentle homogenates of hair bulbs and melanoma results in significantly increased T1 activity within polyacrylamide gels. Similar treatment of 100,000g supernatants results in a slight increase in T1 activity. Addition of Triton-X or DOC to 35,000g supernatants of hair bulb and melanoma homogenates followed by centrifugation at 100,000g results in a marked enhancement of T1 when the latter supernatants are treated with trypsin. In the absence of trypsin treatment, T1 activity is comparable to nondetergent-treated controls. A slow-moving dopa-reactive band (Ts) is found in electropherograms of the nontrypsinized 100,000g supernants of detergent-treated 35,000g supernatants. It is absent in those treated with trypsin. The slow-moving enzyme appears to give rise to T1 molecules when eluted from acrylamide gels and even to a greater extent when elution is combined with trypsin treatment prior to reelectrophoresis. In mammals, tyrosinase apparently is not derived by a proteolytic activation of protyrosinase.
在C57BL小鼠和哈丁 - 帕西小鼠黑色素瘤的色素沉着皮下组织（毛球）中，酪氨酸酶的T1变体以颗粒形式和可溶或易溶解形式存在。用胰蛋白酶处理含有毛球和黑色素瘤温和匀浆微粒体（小颗粒）部分的35,000g上清液，会使聚丙烯酰胺凝胶内的T1活性显著增加。对100,000g上清液进行类似处理，T1活性略有增加。向毛球和黑色素瘤匀浆的35,000g上清液中添加Triton - X或脱氧胆酸钠（DOC），然后以100,000g离心，当用胰蛋白酶处理后者上清液时，T1会显著增强。在没有胰蛋白酶处理的情况下，T1活性与未用去污剂处理的对照相当。在经去污剂处理的35,000g上清液的未用胰蛋白酶处理的100,000g上清液的电泳图中发现了一条迁移缓慢的多巴反应带（Ts）。用胰蛋白酶处理的样品中没有这条带。当从丙烯酰胺凝胶中洗脱时，这条迁移缓慢的酶似乎会产生T1分子，在重新电泳之前，与胰蛋白酶处理结合洗脱时，产生的T1分子更多。在哺乳动物中，酪氨酸酶显然不是由原酪氨酸酶的蛋白水解激活产生的。
