Kidson S H, Fabian B C
J Exp Zool. 1981 Jan;215(1):91-7. doi: 10.1002/jez.1402150111.
The tyrosinase activity of Himalayan mouse skin homogenates was measured over a range of temperatures using two sensitive radiometric assay--namely, (1) the measurement of 14C-tyrosine incorporation into melanin, and (2) the measurement of 3HOH released as a by-product of 3H-tyrosine hydroxylation. Results show that Himalayan tyrosinase is maximally active at temperatures well below normal body temperature (15 degree C to 25 degree C). These results are in support of Danneel's visual observations ('41) that "ferment" activity of Himalayan rabbit skin is absent at temperatures above 25 degree C. Further results suggest the presence of a tyrosinase inhibitor in Himalayan mouse skin. First, removal of a low molecular weight fraction from Himalayan skin homogenates resulted in an increase in tyrosinase activity. Second, recombination of the low molecular weight fraction to the homogenate from which it was originally separated resulted in a decrease in tyrosinase activity when assayed at 37 degree C, but no decrease when assayed at 25 degree C. It is proposed that at the normal body temperature of 37 degree C, tyrosinase from Himalayan skin is strongly bound to an inhibitor. At lower body temperatures, the affinity of the enzyme for the inhibitor decreases, thus allowing the synthesis of melanin to increase. This change in affinity of the enzyme for the inhibitor could be regulated by temperature-induced conformational changes in either the enzyme or the inhibitor or both.
利用两种灵敏的放射性测定法,在一系列温度范围内测量了喜马拉雅小鼠皮肤匀浆的酪氨酸酶活性。这两种方法分别是:(1)测量14C - 酪氨酸掺入黑色素的量;(2)测量作为3H - 酪氨酸羟基化副产物释放的3HOH的量。结果表明,喜马拉雅酪氨酸酶在远低于正常体温(15摄氏度至25摄氏度)的温度下活性最高。这些结果支持了丹内尔(1941年)的视觉观察结果,即喜马拉雅兔皮肤在25摄氏度以上不存在“发酵”活性。进一步的结果表明喜马拉雅小鼠皮肤中存在酪氨酸酶抑制剂。首先,从喜马拉雅皮肤匀浆中去除低分子量部分会导致酪氨酸酶活性增加。其次,将低分子量部分重新组合到其最初分离出的匀浆中,在37摄氏度下测定时酪氨酸酶活性降低,但在25摄氏度下测定时则没有降低。有人提出,在37摄氏度的正常体温下,喜马拉雅皮肤中的酪氨酸酶与一种抑制剂紧密结合。在较低体温下,酶对抑制剂的亲和力降低,从而使黑色素的合成增加。酶对抑制剂亲和力的这种变化可能是由酶或抑制剂或两者的温度诱导构象变化所调节的。