Mapping antibody-binding domains of the major outer surface membrane protein (OspA) of Borrelia burgdorferi.
The major outer surface membrane protein of Borrelia burgdorferi, OspA, is one of several antigens recognized by sera from some patients in the chronic phase of Lyme borreliosis. We have expressed the OspA open reading frame in Escherichia coli and generated a series of deletion constructs of the gene and expressed them as trpE fusion proteins in E. coli. These constructs were used to identify antibody-binding sites of both rabbit antiserum and mouse monoclonal antibodies (MAbs) directed against OspA. All antibodies tested failed to bind to a fusion protein containing the first 61 amino acids of OspA, suggesting that the amino-terminal domain of OspA is unexposed to the cell surface. The binding site for one MAb, 184.1, was identified in a region centered around amino acid 61, while the binding site for MAb 105.5 was identified in a region centered around amino acids 214 to 217. Sera from two patients which were reactive to OspA identified distinct epitopes that lie between those recognized by our MAbs.
Schubach
W H
WH
Department of Medicine, State University of New York, Stony Brook 11794.
Mudri
S
S
Dattwyler
R J
RJ
Luft
B J
BJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D001666:Binding Sites, Antibody / Q000276:immunology*; D015748:Borrelia burgdorferi Group / Q000235:genetics / Q000276:immunology*; D002872:Chromosome Deletion; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000235:genetics; D005798:Genes, Bacterial / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D008193:Lyme Disease / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins