Characterization of a putative receptor protein for bovine viral diarrhea virus.
In a previous communication, we reported a 50-kDa cell surface protein from Madin-Darby bovine kidney (MDBK) cells as a putative receptor for bovine viral diarrhea virus (BVDV). The present study delineates further characterization of the receptor protein. Protease treatment of cultured MDBK cells adversely affected the receptor, thus abolishing the binding of anti-D89 (BVDV anti-idiotypes) to the cells. However, pretreatment of the cells with either phospholipases or glycosidases did not significantly change the anti-D89 binding to the cells. Additionally, pretreatment of cell monolayers with proteases decreased BVDV attachment and replication in the cells. These results suggested that the receptor for BVDV is a protein in nature, and glycosylation and phosphorylation may not play a direct role in BVDV attachment to cells. The BVDV receptor gradually regenerated on the cell surface after the protease-treated cells were cultured in normal growth medium. Regeneration of the BVDV receptor to a normal level took about 4 h as indicated by flow cytometric analysis and this process was inhibited in the presence of cycloheximide, a protein synthesis inhibitor. The 50-kDa receptor protein purified by electro-elution inhibited BVDV infection in a plaque reduction assay. It also inhibited anti-D89 binding to cells as analyzed by flow cytometry. These data demonstrated the nature of the 50-kDa protein as a specific receptor for BVDV.
Xue
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Kansas State University, Manhattan 66506, USA.
Zhang
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Minocha
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HC
D000818:Animals; D001912:Bovine Virus Diarrhea-Mucosal Disease / Q000821:virology*; D002417:Cattle; D002460:Cell Line; D010450:Endopeptidases / Q000494:pharmacology; D007668:Kidney; D007700:Kinetics; D008565:Membrane Proteins / Q000302:isolation & purification; D010576:Pestivirus / Q000502:physiology*; D011991:Receptors, Virus / Q000096:biosynthesis / Q000302:isolation & purification / Q000502:physiology*; D013379:Substrate Specificity; D010948:Viral Plaque Assay; D014779:Virus Replication
D008565:Membrane Proteins; D011991:Receptors, Virus; D010450:Endopeptidases