Intracellular maturation of mumps virus hemagglutinin-neuraminidase glycoprotein: conformational changes detected with monoclonal antibodies.
Monoclonal antibodies elicited by immunization with mumps virus glycoproteins were selected with either native or chymotrypsin-treated mumps virus in an enzyme-linked immunosorbent assay. Group I antibodies which preferentially recognized chymotrypsin-treated virus failed to recognize native mumps virus hemagglutinin-neuraminidase (HN). They did react with sodium dodecyl sulfate-denatured HN and the HN chymotryptic fragments HNc2' (molecular weight, 41,000) and HNc1 (molecular weight, 32,000) after transfer to nitrocellulose paper. In contrast, group II antibodies, which preferentially recognized native virus in the enzyme-linked immunosorbent assay, reacted with native HN but failed to bind HN after sodium dodecyl sulfate denaturation. These two groups of monoclonal antibodies were used to define the maturation pathway of the mumps virus HN in infected cells. The HN initially appeared as a 76,000-molecular-weight polypeptide and was recognized only by group I antibodies. A truncated form of HN, HNT (molecular weight, 63,000), was synthesized in the presence of tunicamycin and was also recognized only by group I antibodies. The 76,000-molecular-weight HN was rapidly converted to a 74,000-molecular-weight polypeptide; this form of HN was recognized only by group II antibodies. The oligosaccharide side chains were modified, and intermolecular disulfide bonds were formed as HN was transported to the cell surface. The disulfide-linked oligomers of HN were direct precursors of the HN found in mature virus.
D000911:Antibodies, Monoclonal; D002451:Cell Compartmentation; D002460:Cell Line; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism; D015707:HN Protein; D006389:Hemagglutinins, Viral / Q000276:immunology / Q000378:metabolism*; D006596:Hexosaminidases / Q000378:metabolism; D008565:Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D009109:Mumps virus / Q000378:metabolism*; D009439:Neuraminidase / Q000276:immunology / Q000378:metabolism*; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D014415:Tunicamycin / Q000494:pharmacology; D014759:Viral Envelope Proteins / Q000276:immunology / Q000378:metabolism*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D015707:HN Protein; D006389:Hemagglutinins, Viral; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014415:Tunicamycin; D006596:Hexosaminidases; D009439:Neuraminidase