Krishnamoorthy Ganesh Kumar, Alluvada Prashanth, Hameed Mohammed Sherieff Shahul, Kwa Timothy, Krishnamoorthy Janarthanan
Curtiss-Wright Avionics and Electronics, Dublin 14, Ireland.
Department of Mechanical Engineering, Jimma Institute of Technology, Jimma University, Ethiopia.
Biochem Biophys Rep. 2019 Dec 17;21:100712. doi: 10.1016/j.bbrep.2019.100712. eCollection 2020 Mar.
Biophysical techniques such as isothermal titration calorimetry (ITC) and surface plasmon resonance (SPR) are routinely used to ascertain the global binding mechanisms of protein-protein or protein-ligand interaction. Recently, Dumas etal, have explicitly modelled the instrument response of the ligand dilution and analysed the ITC thermogram to obtain kinetic rate constants. Adopting a similar approach, we have integrated the dynamic instrument response with the binding mechanism to simulate the ITC profiles of equivalent and independent binding sites, equivalent and sequential binding sites and aggregating systems. The results were benchmarked against the standard commercial software Origin-ITC. Further, the experimental ITC chromatograms of 2'-CMP + RNASE and BH3I-1 + hBCL interactions were analysed and shown to be comparable with that of the conventional analysis. Dynamic approach was applied to simulate the SPR profiles of a two-state model, and could reproduce the experimental profile accurately.
等温滴定量热法（ITC）和表面等离子体共振（SPR）等生物物理技术通常用于确定蛋白质-蛋白质或蛋白质-配体相互作用的整体结合机制。最近，杜马斯等人明确模拟了配体稀释的仪器响应，并分析了ITC热谱图以获得动力学速率常数。采用类似的方法，我们将动态仪器响应与结合机制相结合，以模拟等效独立结合位点、等效顺序结合位点和聚集系统的ITC图谱。结果以标准商业软件Origin-ITC为基准进行了验证。此外，对2'-CMP + RNASE和BH3I-1 + hBCL相互作用的实验ITC色谱图进行了分析，结果表明与传统分析结果具有可比性。应用动态方法模拟了双态模型的SPR图谱，能够准确再现实验图谱。
