Study on the thermodynamics and regulation of amyloid formation using calorimetry

为了研究蛋白质错误折叠诱导聚集的热力学，建立一种新的测定方法，对几种淀粉样蛋白和多肽的淀粉样蛋白的形成进行了等温滴定热法(ITC)的研究。我揭示了淀粉样蛋白的形成在滞后时间后产生了放热，并使用内部算法获得了热力学参数，如热变化。我进一步要求阐明淀粉样蛋白形成的动力学特性，并能够计算出淀粉样蛋白纤颤的成核和伸长的速率常数。利用这些参数构建的能量图谱揭示了b2m淀粉样蛋白的多态发生。基于ITC的检测为研究蛋白质聚集的热力学、动力学和机制提供了强有力的方法。

In order to study thermodynamics of protein misfolding-induced aggregation and establish a new assay, isothermal titration calorimetry (ITC) was performed on amyloidogenesis of several amyloidogenic proteins and peptides. I revealed that amyloid formation resulited in exothermic heat after the lag time, and obtained thermodynamic parameters such as the change in enthalpy using in-house algorithms. I further challenged to elucidate the kinetic property of amyloid formation, and could calculate the rate constant of nucleation and elongation of amyloid fibrillation. Energy landscape built using these parameters revealed the polymorphic amyloidogenesis of b2m.ITC-based assay provides a powerful method for the study of the thermodynamics, kinetics, and mechanism of protein aggregation.