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J Gen Virol 75 (1994), 1163-1167; DOI 10.1099/0022-1317-75-5-1163
© 1994 Society for General Microbiology

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Expression of the Sendai Virus Fusion Protein in Insect Cells and Characterization of its Post-translational Modifications

Evgeni Ponimaskin, Michael Veit and Michael F. G. Schmidt

Institut für Immunologie und Molekularbiologie, Fachbereich Veterinärmedizin, Freie Universität Berlin, Königin-Luise-Strasse 49, 14195 Berlin, Germany

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 Mr 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.

Received 22 September 1993; accepted 30 November 1993.


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