Expression of the Sendai virus fusion protein in insect cells and characterization of its post-translational modifications.
The fusion (F) protein of Sendai virus was expressed with a baculovirus system in insect cells. The F protein is synthesized in its uncleaved form F0, which appears in SDS-containing gels as two bands with M(r) values of 66K and 64K. Digestion of the F protein with endoglycosidase H revealed that the 66K species contains high mannose-type carbohydrates, whereas partially processed oligosaccharides are attached to the 64K species. Pulse-chase experiments showed that the F protein is initially synthesized as its 66K form. After 1 h of chase this precursor glycoprotein is partially converted to the 64K species and exposed at the cell surface. Other modifications of the F protein in insect cells are the formation of intramolecular disulphide linkages and oligomerization. However, processing and plasma membrane transport of the F protein in insect cells are incomplete and retarded compared to the F protein synthesized in Sendai virus-infected mammalian cells.
Ponimaskin
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E
Institut für Immunologie und Molekularbiologie, Fachbereich Veterinärmedizin, Freie Universität Berlin, Germany.
Veit
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Schmidt
M F
MF
D000818:Animals; D001692:Biological Transport; D002451:Cell Compartmentation; D002478:Cells, Cultured; D006031:Glycosylation; D006596:Hexosaminidases / Q000378:metabolism; D009036:Moths / Q000166:cytology; D017924:Nucleopolyhedroviruses / Q000235:genetics; D009844:Oligosaccharides / Q000378:metabolism; D010222:Parainfluenza Virus 1, Human / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis; D014760:Viral Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
D009844:Oligosaccharides; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins; D006596:Hexosaminidases