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Glycosylation pathways of two major Epstein-Barr virus membrane antigens
Abstract:
Monoclonal antibodies to the Epstein-Barr virus (EBV) membrane antigens (MA) were used to delineate the sequence of events occurring during MA maturation. Immunoprecipitation experiments performed with B95-8 virus infected 1605L cells, pulse labeled in the presence of tunicamycin or 2-deoxyglucose, suggest that the nonglycosylated forms of the mature 236,000 (236K) and 86K MA have molecular weights of 149K and 69K, respectively. Monensin treatment results in the accumulation of a 189K antigen which is converted to the mature 236K MA after a 2 hr chase in the absence of monensin. Experiments with monensin also suggest that an 84K intermediate is involved in the formation of the mature 86K MA. The mature 236K MA is resistant to endoglycosidase H while the 189K intermediate and the mature 86K MA are partially sensitive to this enzyme. The mature 236K and 86K MA and the 189K intermediate are resistant to endoglycosidase D.
Insights
This study used monoclonal antibodies to map Epstein-Barr virus (EBV) membrane antigen (MA) maturation. Researchers identified nonglycosylated forms and intermediates, revealing key steps in EBV MA development.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Epstein-Barr virus (EBV) is a human herpesvirus implicated in various cancers.
- Understanding EBV membrane antigen (MA) maturation is crucial for viral pathogenesis research.
Purpose of the Study:
- To delineate the sequence of events during EBV MA maturation.
- To characterize the molecular weights and glycosylation states of EBV MA intermediates.
Main Methods:
- Utilized monoclonal antibodies against EBV MA.
- Performed immunoprecipitation experiments on infected cells.
- Employed pulse-labeling with tunicamycin and 2-deoxyglucose.
- Investigated the effects of monensin treatment.
- Assessed enzyme sensitivity to endoglycosidase H and D.
Main Results:
- Identified nonglycosylated forms of mature 236K and 86K MA at 149K and 69K, respectively.
- Monensin treatment led to accumulation of a 189K intermediate, converting to 236K MA.
- An 84K intermediate was implicated in 86K MA formation.
- Mature 236K MA was resistant to endoglycosidase H; 189K and 86K MA were partially sensitive.
- All studied antigens were resistant to endoglycosidase D.
Conclusions:
- Elucidated the sequential steps in EBV MA maturation.
- Characterized key nonglycosylated intermediates and their molecular weights.
- Demonstrated differential glycosylation patterns influencing MA processing.