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

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Abstract

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.

Original languageEnglish
Pages (from-to)1892-1901
Number of pages10
JournalBiophysical Journal
Volume96
Issue number5
DOIs
Publication statusPublished - 2009 Jan 1

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SARS Virus
Nucleocapsid Proteins
Differential Scanning Calorimetry
Circular Dichroism
Hot Temperature
Sodium Acetate
Transition Temperature
Thermodynamics
Sodium Chloride
Glycine
Spectrum Analysis
Buffers
Magnetic Resonance Spectroscopy
RNA-Binding Motifs

ASJC Scopus subject areas

  • Biophysics

Cite this

Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein : Experiments and numerical simulations. / Fang, Huey Jen; Chen, Yong Zhong; Mai, Suan Li; Wu, Ming Chya; Chang, Chun Ling; Chang, Chung Ke; Hsu, Yen Lan; Huang, Tai Huang; Chen, Hueih Min; Tsong, Tian Yow; Hu, Chin Kun.

In: Biophysical Journal, Vol. 96, No. 5, 01.01.2009, p. 1892-1901.

Research output: Contribution to journalArticle

Fang, HJ, Chen, YZ, Mai, SL, Wu, MC, Chang, CL, Chang, CK, Hsu, YL, Huang, TH, Chen, HM, Tsong, TY & Hu, CK 2009, 'Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein: Experiments and numerical simulations', Biophysical Journal, vol. 96, no. 5, pp. 1892-1901. https://doi.org/10.1016/j.bpj.2008.10.045
Fang, Huey Jen ; Chen, Yong Zhong ; Mai, Suan Li ; Wu, Ming Chya ; Chang, Chun Ling ; Chang, Chung Ke ; Hsu, Yen Lan ; Huang, Tai Huang ; Chen, Hueih Min ; Tsong, Tian Yow ; Hu, Chin Kun. / Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein : Experiments and numerical simulations. In: Biophysical Journal. 2009 ; Vol. 96, No. 5. pp. 1892-1901.
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