Lu Rui, Li Wen-Wei, Katzir Abraham, Raichlin Yosef, Yu Han-Qing, Mizaikoff Boris
Department of Chemistry, University of Science and Technology of China, Hefei 230026, P.R. China.
Analyst. 2015 Feb 7;140(3):765-70. doi: 10.1039/c4an01495b.
Attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy using a special waveguide based on a silver halide fiber was used for probing the heat-induced secondary structure and conformation changes of bovine serum albumin (BSA). From the secondary derivative and the curve fitting of the obtained ATR-FTIR spectra, the changes of the BSA secondary structure with temperature were clearly identified. Two different thermal denaturation temperature ranges (i.e., 50-52 and 80-82 °C, at which a change of the protein structure occurred) were determined, while only one denaturation temperature was previously identified via classical FTIR measurements. Additionally, taking advantage of two-dimensional correlation spectroscopy more detailed information on changes of the protein secondary structure was revealed. The developed method facilitates in situ, sensitive, and more in-depth probing of protein secondary structures, which represents a significant advancement compared to conventional characterization methods.
利用基于卤化银光纤的特殊波导的衰减全反射傅里叶变换红外（ATR-FTIR）光谱法来探测牛血清白蛋白（BSA）热诱导的二级结构和构象变化。通过对获得的ATR-FTIR光谱进行二阶导数和曲线拟合，清楚地确定了BSA二级结构随温度的变化。确定了两个不同的热变性温度范围（即50-52和80-82°C，此时蛋白质结构发生变化），而之前通过经典FTIR测量仅确定了一个变性温度。此外，利用二维相关光谱揭示了关于蛋白质二级结构变化的更详细信息。所开发的方法有助于对蛋白质二级结构进行原位、灵敏且更深入的探测，与传统表征方法相比，这是一个重大进步。
