| 三种测定蛋白质热稳定性方法的比较 |
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| 引用本文： | 周翠燕,俞敏达,李文奇. 三种测定蛋白质热稳定性方法的比较[J]. 分析测试技术与仪器, 2021, 27(4): 252-259. DOI: 10.16495/j.1006-3757.2021.04.004 |
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| 作者姓名： | 周翠燕 俞敏达 李文奇 |
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| 作者单位： | 清华大学 生命科学与医学研究院,北京 100084;国家蛋白质科学研究(北京)设施清华基地,北京 100084;清华大学 生命科学学院,北京 100084;清华大学 结构生物学高精尖创新中心,北京 100084;北京市第四中学国际校区,北京 100032 |
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| 基金项目： | 北京市结构生物学高精尖创新中心资助项目20151551402北京市生物结构前沿研究中心资助项目20181551183 |
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| 摘 要： | 蛋白质的热稳定性一般使用热变性中点温度(melting temperature，Tm)来表示，即蛋白质解折叠50%时的温度. 测定蛋白质Tm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Tm值进行测定. 结果发现，三种方法测得的Tm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Tm值.
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| 关 键 词： | 蛋白质热稳定性 热变性中点温度 差示扫描量热 圆二色谱 差示扫描荧光 |
| 收稿时间： | 2021-10-12 |
Comparation of Three Measuring Methods for Thermodynamic Stability of Protein |
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| Affiliation: | 1.School of Biomedicine in Tsinghua University, Beijing 100084, China2.National Protein Science Facility(Beijing), Tsinghua University, Beijing 100084, China3.School of Life Sciences, Tsinghua University, Beijing 100084, China4.Beijing Advanced Innovation Center for Structural Biology, Beijng 100084, China5.Beijing NO.4 High School International Campus, Beijng 100032, China |
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| Abstract: | Thermodynamic stability describes the tendency of protein unfolding, generally showed by the melting temperature (Tm) 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 fluorimetry (DSF) were used to measure the Tm values of five proteins with different structural feature. The Tm values of different proteins gained using DSC, CD and DSF methods were compared. In summary, the Tm 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 Tm values. |
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