Overproduction, purification, and biochemical characterization of the dual specificity H1 protein phosphatase encoded by variola major virus.
Smallpox, a highly contagious infectious disease caused by the variola major virus, has an overall mortality rate of about 30%. Because there currently is no specific treatment for smallpox, and the only prevention is vaccination, there is an urgent need for the development of effective antiviral drugs. The dual specificity protein phosphatase encoded by the smallpox virus (H1) is essential for the production of infectious viral particles, making it a promising molecular target for antiviral therapeutics. Here, we report the molecular cloning, overproduction, purification, and initial biochemical characterization of H1 phosphatase, thereby paving the way for the discovery of small molecule inhibitors.
Tropea
Joseph E
JE
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD, USA.
Phan
Jason
J
Waugh
David S
DS
D000438:Alcohols / Q000494:pharmacology; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D009596:Nitrophenols / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D011092:Polyethylene Glycols / Q000494:pharmacology; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D012901:Variola virus / Q000235:genetics*; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*
D000438:Alcohols; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D014764:Viral Proteins; C008644:nitrophenylphosphate; D011092:Polyethylene Glycols; D017027:Protein Tyrosine Phosphatases; D004121:Dimethyl Sulfoxide