Thermostability of the N-Terminal RNA-Binding Domain of the SARS-CoV Nucleocapsid Protein: Experiments and Numerical Simulations

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• 作者：Huey-Jen Fang ; Yong-Zhong Chen ; Mai Suan Li ; Ming-Chya Wu ; Chun-Ling Chang ; Chung-ke Chang ; Yen-lan Hsu ; Tai-huang Huang ; Hueih-Min Chen ; Tian-Yow Tsong ; Chin-Kun Hu
• 刊名：Biophysical Journal
• 出版年：2009
• 出版时间：4 March 2009
• 年：2009
• 卷：96
• 期：5
• 页码：1892-1901
• 全文大小：1279 K

文摘

Differential scanning calorimetry, circular dichroism spectroscopy, nuclear magnetic resonance spectroscopy, and numerical simulations were used to study the thermostability of the N-terminal RNA-binding domain (RBD) of the SARS-CoV nucleocapsid protein. The transition temperature of the RBD in a mixing buffer, composed of glycine, sodium acetate, and sodium phosphate with 100 mM sodium chloride, at pH 6.8, determined by differential scanning calorimetry and circular dichroism, is 48.74°C. Experimental results showed that the thermal-induced unfolding-folding transition of the RBD follows a two-state model with a reversibility >90 % . Using a simple Gō-like model and Langevin dynamics we have shown that, in agreement with our experiments, the folding of the RBD is two-state. Theoretical estimates of thermodynamic quantities are in reasonable agreement with the experiments. Folding and thermal unfolding pathways of the RBD also were experimentally and numerically studied in detail. It was shown that the strand β₁ from the N-terminal folds last and unfolds first, while the remaining β-strands fold/unfold cooperatively.