Bujalowski Wlodzimierz, Jezewska Maria J, Bujalowski Paul J
Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA; Department of Obstetrics and Gynecology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA; The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA.
Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA; The Sealy Center for Structural Biology, Sealy Center for Cancer Cell Biology, The University of Texas Medical Branch at Galveston, 301 University Boulevard, Galveston, TX 77555-1053, USA.
Biophys Chem. 2017 Mar;222:7-24. doi: 10.1016/j.bpc.2016.12.006. Epub 2017 Jan 3.
Obtaining a detailed knowledge about energetics of ligand-macromolecule interactions is a prerequisite for elucidation of the nature, behavior, and activities of the formed complexes. The most commonly used methods in characterizing molecular interactions are physico-chemical techniques based mainly on spectroscopic, calorimetric, hydrodynamic, etc., measurements. The major advantage of the physico-chemical methods is that they do not require large quantities of material and, if performed carefully, do not perturb examined reactions. Applications of several different physico-chemical approaches, commonly encountered in analyses of biochemical interactions, are here reviewed and discussed, using examples of simple binding reactions. It is stressed that without determination of the relationship between the measured signal and the total average degree of binding, the performed analysis of a single physico-chemical titration curve may provide only fitting parameters, instead of meaningful interaction parameters, already for the binding systems with only two ligand molecules. Some possible pitfalls in the analyses of single titration curves are discussed.
获取有关配体-大分子相互作用能量学的详细知识是阐明所形成复合物的性质、行为和活性的前提条件。表征分子相互作用最常用的方法是主要基于光谱、量热、流体动力学等测量的物理化学技术。物理化学方法的主要优点是它们不需要大量材料，并且如果操作仔细，不会干扰所研究的反应。本文以简单结合反应为例，对生化相互作用分析中常见的几种不同物理化学方法的应用进行了综述和讨论。需要强调的是，对于仅具有两个配体分子的结合系统，如果不确定测量信号与总平均结合度之间的关系，那么对单个物理化学滴定曲线进行的分析可能仅提供拟合参数，而不是有意义的相互作用参数。文中还讨论了单滴定曲线分析中一些可能存在的陷阱。
