Neutralizing antigenic domains on bovine coronavirus gp100/E2 were mapped to fragments of this protein by proteolytic cleavage and fragment analysis. The procedure involved analysis of fragments generated after incubation of E2–monoclonal antibody complexes with various proteases. The smallest antibody-bound fragments obtained were a 50K fragment following Staphylococcus aureus V8 protease and submaxillary protease digestion, and a 37K fragment following trypsin digestion. Trypsin also produced a transient antibody-bound 50K fragment. A 40K fragment which was not bound by antibody was observed following digestions with all three proteases. The 50K fragments generated by V8, submaxillary protease and trypsin comigrated on gels and displayed the same altered mobility under non-reducing conditions, suggesting identity of these fragments and indicating the presence of disulphide linkages in these fragments. The 40K fragments generated by these three enzymes also comigrated and displayed the same altered mobility under non-reducing conditions. The 37K trypsin fragment contained both neutralizing domains, A and B.
BinnsM. M.,
BoursnellM. E. G.,
CavanaghD.,
PapplnD. J. C.,
BrownT. D. K.1985; Cloning and sequencing of the gene encoding the spike protein of the coronavirus IBV. Journal of General Virology 66:719–726
CavanaghD.,
DavisP. J.,
PappinD. J. C.,
BinnsM. M.,
BoursnellM. E. G.,
BrownT. D. K.1986; Coronavirus IBV: partial amino terminal sequencing of spike polypeptide S2 identifies the sequence Arg-Arg-Phe-Arg-Arg at the cleavage site of the spike precursor propolypeptide of the IBV strains Beaudette and M41. Virus Research 4:133–143
ClevelandD. W.,
FischerS. G.,
KirschnerM. W.,
LaemmliU. K.1977; Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. Journal of Biological Chemistry 252:1102–1106
De GrootR. J.,
MaduroJ.,
LenstraJ. A.,
HorzinekM. C.,
Van Der ZeijstB. A. M.,
SpaanW. J. M.1987; cDNA cloning and sequence analysis of the gene encoding the peplomer protein of feline infectious peritonitis virus. Journal of General Virology 68:2639–2646
DelmasB.,
GelfiJ.,
LaudeH.1986; Antigenic structure of transmissible gastroenteritis virus. II. Domains in the peplomer glycoprotein. Journal of General Virology 67:1405–1418
DeregtD.,
BabiukL. A.1987; Monoclonal antibodies to bovine coronavirus: characteristics and topographical mapping of neutralizing epitopes on the E2 and E3 glycoproteins. Virology 161:410–420
DeregtD.,
SabaraM.,
BabiukL. A.1987; Structural proteins of bovine coronavirus and their intracellular processing. Journal of General Virology 68:2863–2877
EisenbergR. J.,
LongD.,
PereiraL.,
HamparB.,
ZweigM.,
CohenG. H.1982; Effect of monoclonal antibodies on limited proteolysis of native glycoprotein gD of herpes simplex virus type 1. Journal of Virology 41:478–488
LuytjesW.,
SturmanL. S.,
BredenbeekP. J.,
ChariteJ.,
Van Der ZeijstB. A. M.,
HorzinekM. C.,
SpaanW. J. M.1987; Primary structure of the glycoprotein E2 of coronavirus MHV-A59 and identification of the trypsin cleavage site. Virology 161:479–487
MakinoS.,
FlemingJ. O.,
KeckJ. G.,
StohlmanS. A.,
LaiM. M. C.1987; RNA recombination of coronaviruses: localization of neutralizing epitopes and neuropathogenic determinants on the carboxyl terminus of peplomers. Proceedings of the National Academy of SciencesU.S.A 84:6567–6571
MockettA. P. A.,
CavanaghD.,
BrownT. D. K.1984; Monoclonal antibodies to the S1 spike and membrane proteins of avian infectious bronchitis coronavirus strain Massachusetts M41. Journal of General Virology 65:2281–2286
Niesters H. G. M.,
Bleumink-PluymN. M. C.,
OsterhausA. D. M. E.,
HorzinekM. C.,
Van Der ZeijstB. A. M.1987; Epitopes on the peplomer protein of infectious bronchitis virus strain M41 as defined by monoclonal antibodies. Virology 161:511–519
RasschaertD.,
LaudeH.1987; The predicted primary structure of the peplomer protein E2 of the porcine coronavirus transmissible gastroenteritis virus. Journal of General Virology 68:1883–1890
SchmidtI.,
SkinnerM.,
SiddellS.1987; Nucleotide sequence of the gene encoding the surface projection glycoprotein of coronavirus MHV-JHM. Journal of General Virology 68:47–56
SturmanL. S.,
RicardC. S.,
HolmesK. V.1985; Proteolytic cleavage of the E2 glycoprotein of murine coronavirus: activation of cell-fusing activity of virions by trypsin and separation of two different 90K cleavage fragments. Journal of Virology 56:904–911
TalbotP. J.,
SalmiA. A.,
KnoblerR. L.,
BuchmeierM. J.1984; Topographical mapping of epitopes on the glycoproteins of murine hepatitis virus-4 (JHM): correlation with biological activities. Virology 132:250–260
WegeH.,
DorriesR.,
WegeH.1984; Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes on the peplomer protein (E2). Journal of General Virology 65:1931–1942