Advances in the analysis of isothermal titration calorimetry data for ligand-DNA interactions
Advances in the analysis of isothermal titration calorimetry data for ligand-DNA interactions
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DOI：
10.1016/j.ymeth.2007.01.010
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发表时间：
2007-06-01
期刊：
影响因子：
4.8
通讯作者：
Haq, Ihtshamul
中科院分区：
文献类型：
--
作者：
Buurma, Niklaas J.;Haq, Ihtshamul
等温滴定量热法（ITC）是一种用于研究生物相互作用的成熟技术。ITC的优势在于它直接测量与相互作用相关的焓变化。实验还可以产生结合等温线，允许量化平衡结合常数，因此可以建立几乎完整的热力学曲线。ITC的原理和应用在最近几年已经有了很好的记录，实验上该技术使用简单，在理想情况下数据分析是微不足道的。然而，ITC实验可以被设计成使得可以评估先前不可访问的参数。我们概述了其中的一些进展，包括（1）利用不同的实验条件;（2）低亲和力系统;（3）高亲和力系统和置换分析。此外，我们还提出了一个问题：如果无法使用ITC附带的数据分析中包含的拟合函数来拟合数据，该怎么办？可能生成此类数据的实例包括遵循非1：n结合模式的系统和其中结合与其他事件（例如配体解离）偶联的系统。处理这些数据的模型现在出现在文献中，我们总结了配体-DNA相互作用的研究相关的例子。(c)2007年爱思唯尔公司All rights reserved.
Isothermal titration calorimetry (ITC) is a well established technique for the study of biological interactions. The strength of ITC is that it directly measures, enthalpy changes associated with interactions. Experiments can also yield binding isotherms allowing quantification of equilibrium binding constants, hence an almost complete thermodynamic profile can be established. Principles and application of ITC have been well documented over recent years, experimentally the technique is simple to use and in ideal scenarios data analysis is trivial. However, ITC experiments can be designed such that previously inaccessible parameters can be evaluated. We outline some of these advances, including (1) exploiting different experimental conditions; (2) low affinity systems; (3) high affinity systems and displacement assays. In addition we ask the question: What if data cannot be fit using the fitting functions incorporated in the data-analysis that came with your ITC? Examples where such data might be generated include systems following non 1:n binding patterns and systems where binding is coupled to other events such as ligand dissociation. Models dealing with such data are now appearing in literature and we summarise examples relevant for the study of ligand-DNA interactions. (c) 2007 Elsevier Inc. All rights reserved.