title: Thermostability of the N-terminal RNA-binding domain of the SARS-CoV nucleocapsid protein: experiments and numerical simulations.
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.
authors.affiliation: Institute of Physics, Academia Sinica, Nankang, Taipei, Taiwan.
authors.forename: Huey-Jen
authors.initials: HJ
authors.lastname: Fang
authors.forename: Yong-Zhong
authors.initials: YZ
authors.lastname: Chen
authors.forename: Mai Suan
authors.initials: MS
authors.lastname: Li
authors.forename: Ming-Chya
authors.initials: MC
authors.lastname: Wu
authors.forename: Chun-Ling
authors.initials: CL
authors.lastname: Chang
authors.forename: Chung-ke
authors.initials: CK
authors.lastname: Chang
authors.forename: Yen-lan
authors.initials: YL
authors.lastname: Hsu
authors.forename: Tai-huang
authors.initials: TH
authors.lastname: Huang
authors.forename: Hueih-Min
authors.initials: HM
authors.lastname: Chen
authors.forename: Tian-Yow
authors.initials: TY
authors.lastname: Tsong
authors.forename: Chin-Kun
authors.initials: CK
authors.lastname: Hu
mesh_terms: D000465:Algorithms; D002152:Calorimetry, Differential Scanning; D002942:Circular Dichroism; D003198:Computer Simulation; D000086462:Coronavirus Nucleocapsid Proteins; D008956:Models, Chemical; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D019590:Nucleocapsid Proteins / Q000737:chemistry* / Q000378:metabolism; D017510:Protein Folding; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000737:chemistry*; D013696:Temperature; D013816:Thermodynamics; D044366:Transition Temperature
chemical_list: D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D012313:RNA