Coronavirus JHM contained six major proteins, four of which were glycosylated. The proteins were gp170, gp98, gp65, p60, gp25 and p23. Sac(-) cells infected with JHM shut off host cell protein synthesis, and the synthesis of three major (150K, 60K and 23K) and three minor (65K, 30K and 14K) polypeptides was detected by pulse-labelling with 35S-methionine. Antiserum directed against purified virus proteins specifically immunoprecipitated the three major intracellular species and also the 65K minor species. During a chase period, species 150K and 23K were processed and three new immunoprecipitable species, 170K, 98K and 25K appeared. The intracellular species 170K, 98K, 65K, 60K, 25K and 23K co-electrophoresed with virion proteins.
Two-dimensional gel electrophoresis of infected cell polypeptides showed that the 60K, 23K, 25K and 14K species were relatively basic polypeptides whilst the 98K and 170K were relatively acidic and heterogeneously charged polypeptides. Additionally, a charge-size modification of the 23K species during processing was detected, which was not apparent using one-dimensional gel analysis.
BondC. W.,
LeibowitzJ. L.,
RobbJ. A.1979; Pathogenetic murine coronaviruses. II. Characterization of virus specific proteins of murine coronaviruses JHMV and A59V. Virology 94:371–384
KesslerS. W.1975; Rapid isolation of antigens from cells with a staphylococcal protein A antibody absorbent: parameters of the interaction of antibody-antigen complexes with protein A. Journal of Immunology 115:1617–1624
LaskeyR. A.,
MillsA. D.1975; Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. European Journal of Biochemistry 56:335–341
NagashimaK.,
WegeH.,
MeyermannR.,
Ter MeulenV.1979; Demyelinating encephalomyelitis induced by a long term coronavirus infection in rats. Acta Neuropathologica 45:205–213
O’farrellP. Z.,
GoodmanH. M.,
O’farrellP. H.1977; High resolution two dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
SmithA. E.,
WheelerT.,
GlanvilleN.,
KaariainenL.1974; Translation of Semliki Forest virus 42S RNA in a mouse cell-free system to give virus coat proteins. European Journal of Biochemistry 49:101–110
SturmanL. S.1977; Characterization of a coronavirus. I. Structural proteins: effect of preparative conditions on the migration of protein in polyacrylamide gels. Virology 77:637–649
SturmanL. S.,
HolmesK. V.1977; Characterization of a coronavirus. II. Glycoproteins of the viral envelope: tryptic peptide analysis. Virology 77:650–660