In vitro biochemical and thermodynamic characterization of nucleocapsid protein of SARS.
The major biochemical and thermodynamic features of nucelocapsid protein of SARS coronavirus (SARS_NP) were characterized by use of non-denatured gel electrophoresis, size-exclusion chromatographic and surface plasmon resonance (SPR) techniques. The results showed that SARS_NP existed in vitro as oligomer, more probably dimer, as the basic functional unit. This protein shows its maximum conformational stability near pH 9.0, and it seems that its oligomer dissociation and protein unfolding occur simultaneously. Thermal-induced unfolding for SARS_NP was totally irreversible. Both the thermal and chemical denaturant-induced denaturation analyses showed that oligomeric SARS_NP unfolds and refolds through a two-state model, and the electrostatic interactions among the charge groups of SARS_NP made a significant contribution to its conformational stability.
Luo
Haibin
H
Drug Discovery and Design Center, State Key Lab of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China.
Ye
Fei
F
Sun
Tao
T
Yue
Liduo
L
Peng
Shuying
S
Chen
Jing
J
Li
Guowei
G
Du
Yi
Y
Xie
Youhua
Y
Yang
Yiming
Y
Shen
Jianhua
J
Wang
Yuan
Y
Shen
Xu
X
Jiang
Hualiang
H
D002850:Chromatography, Gel; D002942:Circular Dichroism; D000086462:Coronavirus Nucleocapsid Proteins; D019281:Dimerization; D019791:Guanidine / Q000378:metabolism; D006358:Hot Temperature; D066298:In Vitro Techniques; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D019791:Guanidine