Freiburger Lee A, Auclair Karine, Mittermaier Anthony K
Department of Chemistry, McGill University, 801 Sherbrooke Street West, Montréal, Québec, Canada, H3A 2K6.
Thermochim Acta. 2012 Jan 10;527(1):148-157. doi: 10.1016/j.tca.2011.10.018.
Isothermal titration calorimetry (ITC) can provide detailed information on the thermodynamics of biomolecular interactions in the form of equilibrium constants, , and enthalpy changes, Δ . A powerful application of this technique involves analyzing the temperature dependences of ITC-derived and Δ values to gain insight into thermodynamic linkage between binding and additional equilibria, such as protein folding. We recently developed a general method for global analysis of variable temperature ITC data that significantly improves the accuracy of extracted thermodynamic parameters and requires no prior knowledge of the coupled equilibria. Here we report detailed validation of this method using Monte Carlo simulations and an application to study coupled folding and binding in an aminoglycoside acetyltransferase enzyme.
等温滴定量热法（ITC）能够以平衡常数、 和焓变Δ 的形式提供有关生物分子相互作用热力学的详细信息。该技术的一个强大应用是分析ITC得出的 和Δ 值对温度的依赖性，以深入了解结合与其他平衡（如蛋白质折叠）之间的热力学联系。我们最近开发了一种用于可变温度ITC数据全局分析的通用方法，该方法显著提高了提取的热力学参数的准确性，并且不需要对耦合平衡有先验知识。在此，我们报告使用蒙特卡罗模拟对该方法进行的详细验证以及将其应用于研究氨基糖苷乙酰转移酶中的耦合折叠和结合。
