Abstract
Intracellular processing of measles virus fusion (F) protein was studied by radiolabeling and immunoprecipitation with a monoclonal antibody against F protein. The cleavage of F protein into F1 and F2 subunits was complete after 5 hours of chase during which the growth of oligosaccharide chains on the F2 domain of F protein continued. The addition of terminal sialic acid conferred a strong negative charge on the F2 subunit. F protein expressed on the cell surface was removed by a fungal semi-alkaline protease, providing a method to follow the kinetics of its transport to the cell surface. The transport of the F protein was faster than that of the hemagglutinin (HA) protein. Uncleaved F protein, as well as cleaved subunits became digestible by the protease, indicating that a portion of the F protein reaches the cell surface uncleaved. The treatment of measles virus-infected cells with tunicamycin resulted in the synthesis of unglycosylated HA (65 kilodaltons, Kd) and F (48 Kd) proteins. Unglycosylated F protein was not cleaved into smaller subunits, nor was it transported to the cell surface. Unglycosylated HA protein likewise failed to reach the cell surface.
MeSH Terms
Antibodies, Monoclonal
Biological Transport
Glycosylation
Hemagglutinins, Viral/metabolism
Isoelectric Point
Measles virus/metabolism
Membrane Glycoproteins/metabolism
Molecular Weight
Peptide Hydrolases/metabolism
Protein Processing, Post-Translational
Tunicamycin/pharmacology
Viral Fusion Proteins/metabolism
Chemicals
Antibodies, Monoclonal
Hemagglutinins, Viral
Membrane Glycoproteins
Viral Fusion Proteins
Tunicamycin
Peptide Hydrolases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sato T A
Department of Measles Virus, National Institute of Health, Tokyo, Japan.
Kohama T
Sugiura A
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