The different bivalent cations with the same final concentration were put into the dnaase reacting system for 15 minute at ph5 . 4 , at 41 , then dnaase ' s activity unit was detected 在dna酶的酶促反应体系中加入终浓度相同的二价金属阳离子41下、 phs 4准确反应15分钟后测定酶的活力单位。
In order to investigate the effects of ph , temperature , naf and bivalent cations on the conformation of the phosphatase in solution , we monitored the difference of intrinsic fluorescence of the phosphatase and compared changes of the enzyme activity under those conditions . the tertiary structure loosed and the intensity of fluorescence decreased below ph 6 . 0 . the intensity of fluorescence was lowest at ph 5 . 0 and the tertiary structure was reconstructed as the solution ph was increased 其中在ph5 . 0及以下时,蛋白质的三级结构变得松散,荧光强度下降, ph5 . 0时尤为显著,而当溶液ph高于5 . 0时(酶的最适ph ) ,样品的荧光发射强度明显增加,表明酶蛋白受溶液酸碱度的影响,构象发生部分变化,部分trp残基向疏水环境移动,其三级结构得到恢复。
The fluorescence intensity of the phosph atase increased , followed by a decreasing process , in the temperature region of 40 - 90 c . the enzyme was inhibited by naf significantly and the fluorescence spectra of the protein were influenced obviously by the concentration of naf . the data from our experiments showed that the bivalent cations can modulate the activity of the phosphatase but maybe is independent of the tertiary structure Edta对此种磷酸酣酶也有抑制作用,因此用7种二价阳离子处理磷酸酷酶,发现不同金属二价阳离子对这种磷酸酷酶活性和荧光发射光谱的影响不同,因此二价阳离子调控此酶活性,但可能与荧光光谱所反映的三级结构变化无关。