Characterization of a secreted form of measles virus haemagglutinin expressed from a vaccinia virus recombinant.
The measles virus (MV) haemagglutinin (HA) is a class 2 glycoprotein by means of which the virus particle attaches to the host cell receptor. We have previously expressed this glycoprotein as a vaccinia recombinant virus and have shown that the HA glycoprotein synthesized is indistinguishable from that coded by MV. In the present study, we report that in RK13 cells a soluble form (sHA) of the HA is secreted into the medium. We show by SDS-PAGE and sucrose density gradient centrifugation that the sHA is a dimer and is smaller than the cell-associated form. Using a variety of inhibitors the production of sHA was shown to be a late event, probably occurring at the membrane; only fully glycosylated molecules were found in sHA. Finally, we demonstrate that sHA retains its antigenicity with conformation-dependent MAbs and its receptor recognition function. We conclude that sHA is a valuable tool for use in studies of the structure and function of the MV HA glycoprotein.
Malvoisin
E
E
INSERM 404, Immunité et Vaccination, Institut Pasteur de Lyon, France.
Wild
F
F
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015703:Antigens, CD / Q000378:metabolism; D000956:Antigens, Viral / Q000378:metabolism; D001692:Biological Transport; D002460:Cell Line; D003001:Cloning, Molecular; D000939:Epitopes / Q000032:analysis; D006031:Glycosylation; D006389:Hemagglutinins, Viral / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006596:Hexosaminidases; D008459:Measles virus / Q000737:chemistry / Q000378:metabolism*; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000378:metabolism; D008970:Molecular Weight; D011487:Protein Conformation; D011500:Protein Synthesis Inhibitors; D011991:Receptors, Virus / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D014616:Vaccinia virus / Q000235:genetics*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015703:Antigens, CD; D000956:Antigens, Viral; D000939:Epitopes; D006389:Hemagglutinins, Viral; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011500:Protein Synthesis Inhibitors; D011991:Receptors, Virus; D011994:Recombinant Proteins; C032397:hemagglutinin protein G, measles virus; D006596:Hexosaminidases