title: Expression, purification, and initial structural characterization of nonstructural protein 2B, an integral membrane protein of Dengue-2 virus, in detergent micelles.
abstract: Dengue virus causes serious diseases affecting people in tropical and sub-tropical regions. The nonstructural (NS) protein 2B is an integral membrane protein and important for the regulation of viral protease NS3, which is significant for virus replication. The NS2B-NS3 complex is an important drug target for treating dengue fever. However, little is known about the structure of NS2B in its entirety. Herein, we describe the expression and purification of this integral membrane protein from cell membrane and inclusion bodies of Escherichia coli cells. The initial nuclear magnetic resonance (NMR) and circular dichroism (CD) results indicate that the purified protein adopts alpha-helical structures in LMPG and TDPC micelles.
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), Singapore 138669, Singapore.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Huang
authors.forename: Angela Shuyi
authors.initials: AS
authors.lastname: Chen
authors.forename: Qingxin
authors.initials: Q
authors.lastname: Li
authors.forename: Congbao
authors.initials: C
authors.lastname: Kang
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002942:Circular Dichroism; D003001:Cloning, Molecular; D003716:Dengue Virus / Q000737:chemistry*; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification; D008823:Micelles; D008969:Molecular Sequence Data; D010715:Phosphatidylglycerols / Q000737:chemistry; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D017433:Protein Structure, Secondary; D011720:Pyrazoles / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000737:chemistry; D012995:Solubility; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D000085242:Viral Proteases
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D008823:Micelles; D010715:Phosphatidylglycerols; D011720:Pyrazoles; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C552372:lysomyristoylphosphatidylglycerol; C044904:5-tridecylpyrazole-3-carboxylic acid; C504449:NS3 protease, dengue virus; D000085242:Viral Proteases