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Department of Virology, University of Helsinki, Haartmaninkatu 3, SF-00290 Helsinki 29, Finland
Inkoo virus (a bunyavirus) was grown in BHK-21 cells and labelled with [35S]methionine or [3H]mannose. [35S]Methionine labelled the two envelope glycoproteins G1 (Mr = 125000) and G2 (Mr = 35000), as well as the nucleocapsid protein N (Mr = 25000). Only G1 and G2 were labelled with the sugar precursor. The [3H]mannose-labelled virus was solubilized with detergent and digested with Pronase. The structure of the labelled glycopeptides originating from the mixture of G1 and G2 was studied by degrading the glycans stepwise with specific exo- and endoglycosidases, and by analysing the products by both gel and paper chromatography, as well as lectin-affinity chromatography. Three classes of N-glycosidic glycans were found: complex glycans with the monosaccharide sequence (NeuNAc
Gal
GlcNac
)
2 (Man)3 (GlcNAc)2 (occurrence of fucose was not studied), high mannose-type chains with the average structure (Man)46 (GlcNAc)2, and endoglycosidase H-resistant small glycans which were partly susceptible to mannosidase. These latter types of oligosaccharide chains are a novel finding among virus glycoproteins. The relative ratio of the three types of oligosaccharide chains was roughly 4.6:1:1 respectively. The G1 glycoprotein carried most of the sugar chains, since it contained 85% of the [3H]mannose label. The results are discussed in relation to the site of virus maturation at smooth-surfaced vesicles in the Golgi region.
Keywords: bunyavirus, glycan structure, Inkoo virus, glycoproteins
Present address: European Molecular Biology Laboratory, Postfach 10.2209, D-6900 Heidelberg, Federal Republic of Germany.
Received 3 March 1982;
accepted 28 June 1982.
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