Correlated parameter fit of arrhenius model for thermal denaturation of proteins and cells.
Correlated parameter fit of arrhenius model for thermal denaturation of proteins and cells.
复制标题
DOI：
10.1007/s10439-014-1100-y
复制
发表时间：
2014-12
影响因子：
3.8
通讯作者：
Bischof JC
中科院分区：
文献类型：
--
作者：
Qin Z;Balasubramanian SK;Wolkers WF;Pearce JA;Bischof JC
蛋白质的热变性对于细胞损伤、食品科学和其他生物材料加工至关重要。例如，蛋白质变性与加热引起的细胞死亡密切相关，并且在通常超过50 °C的温度下对癌症和其他疾病的局部热疗法越来越感兴趣。阿伦尼乌斯模型是一种简单但广泛使用的蛋白质变性和细胞损伤模型。为了建立Arkenius模型在50 °C及以上的蛋白质变性的实用性，仔细检查了其对动力学参数（活化能Ea和频率因子A）的灵敏度。我们提出了一个简化的相关参数适合Arkyius模型治疗Ea，作为一个独立的拟合参数，并允许A遵循依赖。相关参数拟合的效用被证明对蛋白质和细胞的热变性从文献作为验证，并在我们的实验室使用FTIR光谱的新的实验测量，以证明这种方法的广泛适用性。最后，我们证明了测量变性的最终温度是重要的，并且改变了动力学。具体而言，在低的最终温度（50°C）下发现较高的Ea和A参数，并且随着最终温度增加到70 °C而降低。该趋势与文献中细胞损伤的Arrhenius参数一致，克隆形成（45 - 50 °C）显著高于膜染料测定（60 - 70 °C）。未来的机会，监测细胞损伤的光谱测量蛋白质变性进行了讨论。
Thermal denaturation of proteins is critical to cell injury, food science and other biomaterial processing. For example protein denaturation correlates strongly with cell death by heating, and is increasingly of interest in focal thermal therapies of cancer and other diseases at temperatures which often exceed 50 °C. The Arrhenius model is a simple yet widely used model for both protein denaturation and cell injury. To establish the utility of the Arrhenius model for protein denaturation at 50 °C and above its sensitivities to the kinetic parameters (activation energy Ea and frequency factor A) were carefully examined. We propose a simplified correlated parameter fit to the Arrhenius model by treating Ea, as an independent fitting parameter and allowing A to follow dependently. The utility of the correlated parameter fit is demonstrated on thermal denaturation of proteins and cells from the literature as a validation, and new experimental measurements in our lab using FTIR spectroscopy to demonstrate broad applicability of this method. Finally, we demonstrate that the end-temperature within which the denaturation is measured is important and changes the kinetics. Specifically, higher Ea and A parameters were found at low end-temperature (50°C) and reduce as end-temperatures increase to 70 °C. This trend is consistent with Arrhenius parameters for cell injury in the literature that are significantly higher for clonogenics (45 – 50 °C) vs. membrane dye assays (60 –70 °C). Future opportunities to monitor cell injury by spectroscopic measurement of protein denaturation are discussed.