Purification and characterization of vaccinia virus structural protein VP8.
A major vaccinia virus core protein, designated VP8, has been purified from virions to homogeneity through DEAE-cellulose, CM-cellulose, and hydroxyapatite chromatography. VP8 migrates as a 25-kDa band in SDS-polyacrylamide gel electrophoresis and sediments as a monomeric species in neutral sucrose gradient centrifugation. This protein is a significant constitutent of the virion, comprising about 6.5% of the total viral polypeptides by mass. Analysis by filter binding and by sucrose gradient centrifugation shows that VP8 binds to double-stranded as well as to single-stranded DNA at low salt concentrations (25 mM NaCl). At higher salt concentrations (100 mM NaCl), the protein binds with a relatively greater affinity to single-stranded DNA. The results from sucrose gradient centrifugation indicate that VP8 probably binds noncooperatively to all structural forms of DNA. The protein is likely to be a component of the viral nucleoprotein complex.
Yang
W P
WP
Department of Biochemistry, School of Medicine, State University of New York, Stony Brook 11794.
Bauer
W R
WR
D001667:Binding, Competitive; D002499:Centrifugation, Density Gradient; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008970:Molecular Weight; D014461:Ultracentrifugation; D014616:Vaccinia virus / Q000032:analysis*; D014758:Viral Core Proteins / Q000302:isolation & purification*
D004279:DNA, Viral; D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D014758:Viral Core Proteins