摘要
蛋白质的热稳定性一般使用热变性中点温度(melting temperature,T_(m))来表示,即蛋白质解折叠50%时的温度.测定蛋白质T_(m)值的常用方法有差示扫描量热法(differential scanning calorimetry,DSC)、圆二色光谱法(circular dichroism,CD)和差示扫描荧光法(differential scanning fluorimetry,DSF)等.为了比较不同方法的检测特点和效果,选择了5种具有不同结构特点的蛋白,分别使用上述方法对其T_(m)值进行测定.结果发现,三种方法测得的T_(m)值整体上较为一致,但也存在一定的差异.研究还发现,结构更复杂的蛋白,并不一定具有更多T_(m)值.
Thermodynamic stability describes the tendency of protein unfolding,generally showed by the melting temperature(T_(m))at which 50%of the protein is unfolded.In order to compare the detection characteristics and effects of different methods,differential scanning calorimetry(DSC),circular dichroism(CD)and differential scanning uorimetry(DSF)were used to measure the T_(m) values of five proteins with different structural feature.The T_(m) values of different proteins gained using DSC,CD and DSF methods were compared.In summary,the T_(m) values of the same protein measured by the three methods were rather consistent,but differences were also observed.Additionally,proteins with complex structures did not show multiple T_(m) values.
作者
周翠燕
俞敏达
李文奇
ZHOU Cui-yan;YU Min-da;LI Wen-qi(School of Biomedicine in Tsinghua University,Beijing 100084,China;National Protein Science Facility(Beijing),Tsinghua University,Beijing 100084,China;School of Life Sciences,Tsinghua University,Beijing 100084,China;Beijing Advanced Innovation Center for Structural Biology,Beijng 100084,China;Beijing NO.4 High School International Campus,Beijng 100032,China)
出处
《分析测试技术与仪器》
CAS
2021年第4期252-259,共8页
Analysis and Testing Technology and Instruments
基金
北京市结构生物学高精尖创新中心资助项目(No.20151551402)
北京市生物结构前沿研究中心资助项目(No.20181551183)。
