Inhibition of African swine fever virus binding and infectivity by purified recombinant virus attachment protein p12.
The African swine fever virus protein p12, involved in virus attachment to the host cell, has an apparent molecular mass of 17 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. We have also identified 12- and 10-kDa forms of the p12 protein in infected Vero cells and found that the mature 17-kDa protein is the only form present in virus particles. The p12 protein has been produced in large amounts in Spodoptera frugiperda insect cells infected with a recombinant baculovirus. A 17-kDa protein that possessed the biological properties of the viral protein was produced, since it bound to susceptible Vero cells and not to receptor-negative L cells, which do not support virus replication. The binding of the baculovirus-expressed protein p12 to Vero cells was specifically blocked by virus particles. In addition, the recombinant protein purified by immunoaffinity chromatography blocked the specific binding of virus particles to susceptible cells and prevented infection, demonstrating that the p12 protein mediates the attachment of virions to specific receptors and indicating that blocking the p12-mediated interaction between African swine fever virus and receptors in Vero cells can inhibit infection. However, although antibodies specific for protein p12 are induced in natural infections and in animals inoculated with inactivated virus or recombinant protein p12, these antisera did not inhibit virus binding to the host cell or neutralize virus infectivity.
Angulo
A
A
Centro de Biología Molecular Severo Ochoa (Consejo Superior de Investigaciones Científicas-UAM), Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
Viñuela
E
E
Alcamí
A
A
D000358:African Swine Fever Virus / Q000378:metabolism / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D007739:L Cells; D051379:Mice; D008969:Molecular Sequence Data; D009036:Moths; D010957:Plasmids; D011817:Rabbits / Q000276:immunology; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013552:Swine; D013556:Swine, Miniature / Q000276:immunology; D014162:Transfection; D014709:Vero Cells; D014764:Viral Proteins / Q000096:biosynthesis / Q000378:metabolism* / Q000494:pharmacology
D000906:Antibodies; D011994:Recombinant Proteins; D014764:Viral Proteins; C074550:protein p12, African swine fever virus