Biochimica et Biophysica Acta (BBA) - General Subjects ( IF 2.8 ) Pub Date : 2020-10-22 , DOI: 10.1016/j.bbagen.2020.129772 Steffen Glöckner , Gerhard Klebe
Background
Thermodynamic and binding kinetic data increasingly support and guide the drug optimization process.
Methods
Because ITC thermograms contain binding thermodynamic and kinetic information, an efficient protocol for the simultaneous extraction of thermodynamic and kinetic data for 1:1 protein ligand reactions from AFFINImeter kinITC in one single experiment are presented.
Results
The effort to apply this protocol requires the same time as for the standard protocol but increases the precision of both thermodynamic and kinetic data.
Conclusions
The protocol enables reliable extraction of both thermodynamic and kinetic data for 1:1 protein-ligand binding reactions with improved precision compared to the ‘standard protocol’.
General significance
Thermodynamic and kinetic data are recorded under exactly the same conditions in solution without any labeling or immobilization from a protein sample that is not 100% active and would otherwise render the extraction of kinetic parameters impossible.
中文翻译:
等温滴定量热法同时测定热力学和动力学数据
背景
热力学和结合动力学数据越来越多地支持和指导药物优化过程。
方法
由于ITC温度记录图包含结合的热力学和动力学信息,因此提出了在一个实验中从AFFINImeter kinITC中同时提取1:1蛋白配体反应的热力学和动力学数据的有效方案。
结果
应用此协议需要付出与标准协议相同的时间,但是增加了热力学和动力学数据的精度。
结论
与“标准规程”相比,该规程能够可靠地提取1:1蛋白-配体结合反应的热力学和动力学数据。
一般意义
在完全相同的条件下,在溶液中记录热力学和动力学数据,而没有来自非100%活性蛋白质样品的任何标记或固定,否则将无法提取动力学参数。