Inhibition of proteolytic cleavage of the hemagglutinin of influenza virus by the calcium-specific ionophore A23187.
At calcium-specific ionophore A23187 concentrations of approximately 0.25 microM [which still allow assembly and release of fowl plague virus (FPV) particles] post-translational proteolytic cleavage of the viral hemagglutinin precursor HA into the fragments HA1 and HA2 is inhibited. The resulting virus particles with uncleaved hemagglutinin, that cannot be obtained under normal conditions, provide a suitable substrate for in vitro assays of the protease sensitivity of the FPV hemagglutinin. Proteolytic activation is accomplished with trypsin. Treatment with cathepsin B at low pH yields aberrant cleavage products suggesting that the cellular cleavage enzyme is not of lysosomal origin. A protease that cleaves the FPV hemagglutinin in the correct place can be detected in lysates of MDBK cells. This enzyme is calcium dependent and has a neutral pH optimum.
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology*; D002417:Cattle; D002460:Cell Line; D006224:Cricetinae; D006386:Hemagglutination Tests; D006801:Humans; D009980:Influenza A virus / Q000187:drug effects / Q000235:genetics*; D007668:Kidney; D013820:Thermolysin / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000032:analysis; D014763:Viral Matrix Proteins; D010948:Viral Plaque Assay; D014779:Virus Replication / Q000187:drug effects
C031540:M-protein, influenza virus; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins; D000001:Calcimycin; D013820:Thermolysin