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

Overview

Journal Biophys J

Publisher Cell Press

Specialty Biophysics

Date 2009 Mar 4

PMID 19254548

Authors

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

Affiliations

Soon will be listed here.

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 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 degrees 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 beta(1) from the N-terminal folds last and unfolds first, while the remaining beta-strands fold/unfold cooperatively.

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