Rochu D, Ducret G, Masson P
Centre de Recherches, Unité d'Enzymologie, BP 87, La Tronche, France.
J Chromatogr A. 1999 Apr 9;838(1-2):157-65. doi: 10.1016/s0021-9673(99)00062-x.
The thermal denaturation process of a model protein, bovine beta-lactoglobulin, was analyzed using capillary zone electrophoresis (CZE). For this purpose, a commercial CE apparatus was improved, allowing efficient control and accurate measurement of the temperature up to 95 degrees C. Under various pH conditions, transition temperature (Tm), enthalpy change (delta H) and entropy change (delta S) associated with the thermal denaturation were determined. Moreover, the technique is unique in its ability to estimate the heat capacity change (delta Cp). This work shows that CZE, performed even when electroosmotic flow occurs, is an innovative approach for determining the stability curves of proteins. Accordingly, CZE is a powerful tool to study protein unfolding/folding quickly and with minimal sample requirements.
利用毛细管区带电泳（CZE）分析了模型蛋白牛β-乳球蛋白的热变性过程。为此，对商用CE仪器进行了改进，使其能够在高达95℃的温度下实现高效控制和精确测量。在不同pH条件下，测定了与热变性相关的转变温度（Tm）、焓变（ΔH）和熵变（ΔS）。此外，该技术在估算热容变化（ΔCp）方面具有独特能力。这项工作表明，即使在存在电渗流的情况下进行的CZE，也是一种确定蛋白质稳定性曲线的创新方法。因此，CZE是一种强大的工具，可快速且以最少的样品需求研究蛋白质的展开/折叠。
