title: Studies on the structures and antigenic properties of rabies virus glycoprotein analogues produced in yeast cells.
abstract: We investigated two forms (designated as yGI and yGII) of rabies virus glycoprotein (G) analogues produced in the G cDNA-transfected yeast cells. Molecular weights of yGI and yGII were estimated as 66 and 56 kDa, respectively, according to their relative mobility in SDS-PAGE. Although being produced in large amounts, yGI was present mostly in insoluble forms and hardly extractable with non-ionic detergents. The yGI reacted with polyclonal anti-G antibodies, but did not react with our conformational epitope-specific anti-G monoclonal antibodies (G-MAbs). No protective immunity was induced by yGI in guinea pigs nor in mice. On the other hand, yGII was Triton-soluble, but was only small in amount (at most 1% of total G proteins) and was shown to lack the cytoplasmic domain. The yGII, however, reacted with the G-MAbs and induced protective immunity in guinea pigs as well. When the G-cDNA was expressed in animal cells in culture, a single form (about 66 kDa) of G protein was produced, which displayed similar behaviors as seen in its reactivity with the MAbs and intracellular distribution as seen in the virus-infected cells. These results suggest that most G protein molecules were not processed normally in yeast cells, resulting in abnormal folding and multimer formation, while only a small fraction were occasionally folded normally to have conformational epitopes but were mostly deprived of the C-terminal portion.
authors.affiliation: Research and Development Department, Chemo-Sero-Therapeutic Research Institute, Kumamoto Prefecture, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Sakamoto
authors.forename: T
authors.initials: T
authors.lastname: Ide
authors.forename: S
authors.initials: S
authors.lastname: Tokiyoshi
authors.forename: J
authors.initials: J
authors.lastname: Nakao
authors.forename: F
authors.initials: F
authors.lastname: Hamada
authors.forename: M
authors.initials: M
authors.lastname: Yamamoto
authors.forename: J A
authors.initials: JA
authors.lastname: Grosby
authors.forename: Y
authors.initials: Y
authors.lastname: Ni
authors.forename: A
authors.initials: A
authors.lastname: Kawai
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis*; D003902:Detergents; D004353:Drug Evaluation, Preclinical; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D005814:Genes, Viral; D005822:Genetic Vectors; D006023:Glycoproteins / Q000276:immunology*; D006031:Glycosylation; D006168:Guinea Pigs; D015151:Immunoblotting; D051379:Mice; D017830:Octoxynol; D011820:Rabies virus / Q000276:immunology*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012995:Solubility; D013329:Structure-Activity Relationship