Retarded processing of influenza virus hemagglutinin in insect cells.
When expressed in Spodoptera frugiperda cells by a baculovirus vector, the hemagglutinin of fowl plague virus has been found to contain palmitic acid in covalent hydroxylamine-sensitive linkage, indicating that these cells have the capacity to acylate foreign proteins at cysteine residues. Centrifugation on sucrose density gradients and immune precipitation with conformation-specific antibodies were used to compare trimerization of the hemagglutinin in insect cells and in fowl plague virus-infected MDCK cells. Trimerization of the hemagglutinin was incomplete in insect cells, and the kinetics of this reaction were about three times slower than in vertebrate cells. Similarly, post-translational proteolytic cleavage occurred in insect cells with a half-time of 90 min, and a substantial fraction of the hemagglutinin persisted in uncleaved form. In contrast, hemagglutinin was almost completely cleaved in MDCK cells, and the half-time of cleavage was only 30 min. The data indicate that in insect cells trimerization and, as a result, the subsequent processing steps of the hemagglutinin, are retarded and less efficient. The possible roles of aberrant glycosylation, acidic milieu, and lack of other influenza virus proteins in hemagglutinin trimerization are discussed.
Kuroda
K
K
Institut für Virologie, Philipps-Universität, Marburg, Germany.
Veit
M
M
Klenk
H D
HD
D000215:Acylation; D000818:Animals; D000911:Antibodies, Monoclonal; D001692:Biological Transport; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005057:Eukaryotic Cells; D005227:Fatty Acids / Q000378:metabolism; D006389:Hemagglutinins, Viral / Q000378:metabolism*; D009980:Influenza A virus / Q000254:growth & development / Q000378:metabolism*; D007700:Kinetics; D009036:Moths / Q000166:cytology; D011108:Polymers; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational
D000911:Antibodies, Monoclonal; D005227:Fatty Acids; D006389:Hemagglutinins, Viral; D011108:Polymers