Epitope model of tick-borne encephalitis virus envelope glycoprotein E: analysis of structural properties, role of carbohydrate side chain, and conformational changes occurring at acidic pH.
A panel of monoclonal antibodies (MAbs) was prepared to analyze the antigenic structure of the tick-borne encephalitis (TBE) virus glycoprotein E. Nineteen different epitopes were identified and characterized with respect to serological specificity, functional activity, structural properties, and topological relationships. Except for 3 isolated epitopes (i1, i2, and i3), these cluster to form three non-overlapping domains termed A, B, and C. The structural properties of epitopes were assessed by analyzing the effect of different treatments (SDS denaturation, reduction and carboxymethylation, performic acid oxidation, exposure to pH 5.0, CNBr, and trypsin cleavage) on the antigenic reactivities of each epitope. Only 3 epitopes of domain A as well as i2 were sensitive to SDS alone, whereas all others were SDS resistant. Reduction and carboxymethylation, however, destroyed the antigenic reactivity of all epitopes of domain B and also that of two SDS-resistant epitopes of domain A, indicating the role of disulfide bridges in stabilizing the conformation of these epitopes. Deglycosylation by N-Glycanase abolished the SDS resistance of domain C, providing evidence of the role of the carbohydrate side chain in stabilizing these epitopes. A conformational change induced by acid pH was revealed by differences in protease (proteinase K) cleavage maps before and after acid pH treatment. The conformational change involved the epitopes of domain A and occurred between pH 6.0 and 5.5 with the the threshold at pH 7.0.
Guirakhoo
F
F
Institute of Virology, University of Vienna, Austria.
Heinz
F X
FX
Kunz
C
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis*; D001667:Binding, Competitive; D015153:Blotting, Western; D002241:Carbohydrates / Q000276:immunology; D002478:Cells, Cultured; D004669:Encephalitis Viruses, Tick-Borne / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006023:Glycoproteins / Q000276:immunology*; D006386:Hemagglutination Tests; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D009500:Neutralization Tests; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D002241:Carbohydrates; D000939:Epitopes; D006023:Glycoproteins; D014759:Viral Envelope Proteins