A virus-neutralizing epitope on the glycoprotein of rabies virus that contains Trp251 is a linear epitope.
We have established a hybridoma producing monoclonal antibody (MAb) against a linear epitope of glycoprotein (G protein) of the RC-HL strain of rabies virus. This MAb15-13 showed almost the same neutralizing activity to all of five rabies fixed strains, including RC-HL, and reacted to the denatured G protein in western blot analysis. To characterize and map this linear epitope, an antigenic variant NR15-13 was selected from RC-HL strain in the presence of neutralizing MAb15-13. The variant reacted with MAb15-13 in an immunofluorescent antibody test but was not neutralized by the antibody and the antibody did not bind to the variant G protein in a Western blot analysis. The variant NR15-13 had an amino acid substitution at position 251 of the G protein, where tryptophan of the parental RC-HL strain was replaced by arginine. Site-directed mutagenesis analysis using the expression system in simian COS7 cells revealed that a single amino acid substitution at 251-tryptophan by arginine on the G protein of the parental RC-HL strain abolished the antigenicity of the epitope for MAb15-13 in western blot analysis, and the replacement of 251-arginine by tryptophan recovered the activity. These results strongly suggest that tryptophan at position 251 on the G protein is essential for construction of the linear epitope against MAb15-13.
Luo
T R
TR
Department of Veterinary Public Health, Faculty of Agriculture, Gifu University, Japan.
Minamoto
N
N
Ito
H
H
Goto
H
H
Hiraga
S
S
Ito
N
N
Sugiyama
M
M
Kinjo
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology*; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology*; D019204:GTP-Binding Proteins / Q000276:immunology; D014644:Genetic Variation; D006023:Glycoproteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed; D009500:Neutralization Tests; D017354:Point Mutation / Q000235:genetics / Q000276:immunology; D011820:Rabies virus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017422:Sequence Analysis, DNA; D014364:Tryptophan / Q000032:analysis / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins; D014364:Tryptophan; D019204:GTP-Binding Proteins