Purified foot-and-mouth disease virus (FMDV) of type O1K was treated with several endopeptidases of differing specificity. The immunizing protein VPThr was cleaved into two detectable fragments by all enzymes except for glutamic acid-specific Staphylococcus aureus V8 protease. The longest fragments were generated by mouse submaxillary gland protease and the shortest by trypsin treatment of the intact virion. Several fragments, including the peptides resulting from the cyanogen bromide (CNBr) cleavage of the isolated protein VPThr, were characterized in terms of their molecular weights, N- and C-terminal amino acids, and ability to induce virus-specific antibodies. The order of the fragments along the protein was determined, and then located on the amino acid sequence of the protein. Two enzyme-sensitive areas of the protein were found on the surface of the virion: between sequence positions 138 and 154 and between portion 200 and the C terminus. Peptides containing these sections were able to induce neutralizing antibodies against the homologous FMDV. When the virus was treated with trypsin or with chymotrypsin, several amino acids between the detectable fragments were lost and the infectivity of the virus was reduced. The infectivity was retained, however, when the enzyme treatment resulted in cleavage of protein with no loss of amino acids or only the cutting away of the C-terminal section. These results suggest that the property of cell attachment is restricted to small regions of the surface of the virus particle.
BachrachH. L.,
MooreD. M.,
McKercherP. D.,
PolatnickJ.1975; Immune and antibody response to isolated capsid protein of foot-and-mouth disease virus. Journal of Immunology 115:1636–1641
BartelingS. J.,
MeloenR. H.,
WagenaarF.,
GielkinsA. L. J.1979; Isolation and characterization of trypsin-resistant O1 variants of foot-and-mouth disease virus. Journal of General Virology 43:383–393
BrownF.,
CartwrightB.,
StewartD. L.1963; The effect of various inactivating agents on the viral and the ribonucleic acid infectivities of foot-and-mouth disease virus and on its attachment to susceptible cells. Journal of General Microbiology 31:179–186
BurroughsJ. N.,
RowlandsD. J.,
SangarD. V.,
TalbotP.,
BrownF.1971; Further evidence for multiple proteins in the foot-and-mouth disease virus particle. Journal of General Virology 13:73–84
CavanaghD.,
SangarD. V.,
RowlandsD. J.,
BrownF.1977; Immunogenic and cell attachment sites of FMDV: further evidence for their location in a single capsid polypeptide. Journal of General Virology 35:149–158
HareP. E.1975; Amino-acid composition by column chromatography. In Protein Sequence Determination pp 204–231 Edited by
NeedlemanS. B.
Berlin, Heidelberg & New York: Springer-Verlag;
KaadenO. R.,
AdamK. H.,
StrohmaierK.1977; Induction of neutralizing antibodies and immunity in vaccinated guinea pigs by cyanogen bromide peptides of VP3 of foot-and-mouth disease virus. Journal of General Virology 34:397–400
KupperH.,
KellerW.,
KurzCh.,
ForssS.,
SchallerH.,
FranzeR.,
StrohmaierK.,
MarquardtO.,
ZaslavskyV. G.,
HofschneiderH.1981; Cloning of cDNA of major antigen of foot-and-mouth disease virus and expression in E. coli. Nature, London 289:555–559
KurzC. H.,
ForssS.,
KupperH.,
StrohmaierK.,
SchallerH.1981; Nucleotide sequence and corresponding amino-acid sequence of gene for the major antigen of foot-and-mouth disease virus. Nucleic Acids Research 9:1919–1931
LaporteJ.,
GrosclaudeJ.,
WantyghemJ.,
BernardS.,
RouzeP.1973; Neutralisation en culture cellulaire du pouvoir infectieux du virus de la fièvre aphteuse par des serums provenant de pores immunises a l’aide d’une protéine virale purifiée. Comptes Rendus Hebdomadaires des Seances de TAcadémie des Sciences, Series D 276:3399
La TorreJ. L.,
GrubmanM. J.,
BaxtB.,
BaChrachH. L.1980; The structural polypeptides of aphthovirus are phospho-proteins. Proceedings of the National Academy of Sciences of the United States of America 77:7444–7447
MathekaH. D.,
BachrachH. L.1975; N-Terminal amino acid sequences in the major capsid proteins of foot-and-mouth disease virus types A, O and C. Journal of Virology 16:1248–1253
MathekaH. D.,
MayrA.,
BoegelK.1962; Die Trypsin-Resistenzprufüng zur Differenzierung kleiner chloroformstabiler Virusarten. Zentrablatt für Bakteriologie, Parasitenkunde, Infektionskrankheiten und Hygiene, Abteilung I Originate 187:137–143
MeloenR. H.,
BriaireJ.1980; A study of the cross-reacting antigens on the intact foot-and-mouth disease virus and its 12S subunits with antisera against the structural proteins. Journal of General Virology 51:107–116
RowlandsD. J.,
SangarD. V.,
BrownF.1971; Relationship of the antigenic structure of foot-and-mouth disease virus to the process of infection. Journal of Virology 13:85–93
StrohmaierK.,
AdamK. H.1975; New results concerning the coat proteins of foot-and-mouth disease virus. Bulletin de I’Office International des Epizooties 83:425–429
StrohmaierK.,
KaadenO. R.,
AdamK. H.,
Wittmann-LieboldB.1978a; Estimation of 140S particles by physical techniques, isolation of immunizing peptides, and determination of some sequences of the coat proteins of foot-and-mouth disease virus. Report of the Session of the Research Group of the Standing Technical Committee of the European Commission for the Control of Foot-and-Mouth Disease Brussels Belgium:
StrohmaierK.,
Wittmann-LieboldB.,
GeisslerA. W.1978b; The N terminal sequence of three coat proteins of foot-and-mouth disease virus. Biochemical and Biophysical Research Communications 85:1640–1645
SwankR. T.,
MunkresK. D.1971; Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Analytical Biochemistry 39:462–477
VollerA.,
BidwellD. E.,
BartlettA.1976; Enzyme immunoassay in diagnostic medicine. Theory and practice. Bulletin of the World Health Organization 53:55–65