海藻糖对人脑型肌酸激酶热变性的保护作用  

Protective Effect of Trehalose on Human Brain-type Creatine Kinase During Thermal Denaturation

作　　者：宋青山[1] 杨江流[1] 贾芳[1] 

机构地区：[1]河套学院医学系,巴彦淖尔015000

出　　处：《中国畜牧兽医》2016年第1期157-163,共7页China Animal Husbandry & Veterinary Medicine

基　　金：内蒙古自治区高等学校科学研究项目(2014NJZY14332)

摘　　要：肌酸激酶(creatine kinase,CK)是研究蛋白质折叠和稳定性的一个理想的模型蛋白。热失活研究结果显示海藻糖能有效防止人脑型肌酸激酶(human brain-type creatine kinase,hBBCK)的热失活和热变性。1.0mol/L海藻糖使hBBCK热失活的半失活温度(Tm)升高4.6℃,活化能由原来的29.7kJ/mol升高到41.1kJ/mol。内源荧光光谱研究结果显示在海藻糖浓度分别为0、0.6、0.8和1.2mol/L时,hBBCK热变性的表观转变温度(T1/2)相应的从43.0℃上升到46.5、47.7和49.9℃。内源荧光和ANS外源荧光光谱研究结果表明海藻糖的疏水作用可能对hBBCK的热稳定性起了重要作用。hBBCK复性动力学试验结果表明海藻糖能有效抑制hBBCK复性过程中聚集体的形成。Creatine kinase（CK）is a good model for studying protein folding and stability.The result of thermal inactivation showed that trehalose could prevent the thermal inactivation and thermal denaturation of human brain-type creatine kinase（hBBCK）.In the presence of 1.0mol/L trehalose,the midpoint temperature of thermal inactivation（Tm）of hBBCK increased by 4.6 ℃,and the activation energy（Ea）for thermal inactivation increased from 29.7to 41.1kJ/mol.Intrinsic fluorescence spectra also showed an increase in the apparent transition temperature（T1/2）of hBBCK from 43.0to 46.5,47.7and 49.9℃in 0,0.6,0.8and 1.2mol/L trehalose,respectively.The results of intrinsic fluorescence spectra and extrinsic fluorescence spectra of ANS showed that the hydrophobic effects of trehalose might play an important role in thermal stability of hBBCK.The result of renaturation kinetics experiment showed that trehalose could effectively block the coagulation of hBBCK during renaturation process.

关 键 词：人脑型肌酸激酶 海藻糖 热变性 

分 类 号：Q67[生物学—生物物理学]