Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein: Experiments and numerical simulations

Huey Jen Fang, Yong Zhong Chen, Suan Li Mai, Ming Chya Wu, Chun Ling Chang, Chung Ke Chang, Yen Lan Hsu, Tai Huang Huang, Hueih Min Chen, Tian Yow Tsong, Chin Kun Hu

研究成果: 雜誌貢獻文章

8 引文 斯高帕斯(Scopus)

摘要

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 SARSCoV 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.

原文英語
頁(從 - 到)1892-1901
頁數10
期刊Biophysical Journal
96
發行號5
DOIs
出版狀態已發佈 - 2009 一月 1

    指紋

ASJC Scopus subject areas

  • Biophysics

引用此

Fang, H. J., Chen, Y. Z., Mai, S. L., Wu, M. C., Chang, C. L., Chang, C. K., Hsu, Y. L., Huang, T. H., Chen, H. M., Tsong, T. Y., & Hu, C. K. (2009). Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein: Experiments and numerical simulations. Biophysical Journal, 96(5), 1892-1901. https://doi.org/10.1016/j.bpj.2008.10.045