海藻糖对人脑型肌酸激酶热变性的保护作用

doi:10.16431/j.cnki.1671-7236.2016.01.023

›› 2016, Vol. 43 ›› Issue (1): 157-163.doi: 10.16431/j.cnki.1671-7236.2016.01.023

海藻糖对人脑型肌酸激酶热变性的保护作用

宋青山, 杨江流, 贾芳

河套学院医学系, 巴彦淖尔 015000

收稿日期:2015-05-22 出版日期:2016-01-20 发布日期:2016-01-23
通讯作者: 贾芳 E-mail:Jiafang198011@163.com
作者简介:宋青山(1972-),男,内蒙古乌兰察布人,学士,讲师,研究方向:生物化学,E-mail:nptt2005@163.com
基金资助:
内蒙古自治区高等学校科学研究项目(2014NJZY14332)

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

SONG Qing-shan, YANG Jiang-liu, JIA Fang

Department of Medicine, Hetao College, Bayannaoer 015000, China

Received:2015-05-22 Online:2016-01-20 Published:2016-01-23

摘要/Abstract

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(T_m)升高4.6℃,活化能由原来的29.7 kJ/mol升高到41.1 kJ/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(T_1/2)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。

关键词: 人脑型肌酸激酶; 海藻糖; 热变性

Abstract: 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 1.0 mol/L trehalose, the midpoint temperature of thermal inactivation(T_m) of hBBCK increased by 4.6℃, and the activation energy(E_a) for thermal inactivation increased from 29.7 to 41.1 kJ/mol.Intrinsic fluorescence spectra also showed an increase in the apparent transition temperature(T_1/2) of hBBCK from 43.0 to 46.5, 47.7 and 49.9℃ in 0, 0.6, 0.8 and 1.2 mol/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.

Key words: human brain-type creatine kinase(hBBCK); trehalose; thermal denaturation

中图分类号:

宋青山, 杨江流, 贾芳. 海藻糖对人脑型肌酸激酶热变性的保护作用[J]. , 2016, 43(1): 157-163.

SONG Qing-shan, YANG Jiang-liu, JIA Fang. Protective Effect of Trehalose on Human Brain-type Creatine Kinase During Thermal Denaturation[J]. , 2016, 43(1): 157-163.

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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

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