Characterization of an anti-Borrelia burgdorferi OspA conformational epitope by limited proteolysis of monoclonal antibody-bound antigen and mass spectrometric peptide mapping.
Lyme borreliosis is a multisystem disorder caused by the spirochete Borrelia burgdorferi that is transmitted to humans by the tick Ixodes dammini. The immune response against the 31 kDa OspA, which is one of the most abundant B. burgdorferi proteins, appears to be critical in preventing infection and tissue inflammation. Detailed knowledge of the immunological and molecular characteristics of the OspA protein is important for the development of reliable diagnostic assays. In this study, we characterized a new conformational epitope present within the middle part of B. burgdorferi OspA. Our approach used enzymatic proteolyses of the immune complex followed by mass spectrometric identification of the peptides bound to the antibody. It appears to be one of the first reports on the characterization of a discontinuous epitope using mass spectrometry.
Legros
V
V
Institut de Biologie Structurale (CEA-CNRS-UJF), Laboratoire de Spectrométrie de Masse des Protéines, Grenoble, France.
Jolivet-Reynaud
C
C
Battail-Poirot
N
N
Saint-Pierre
C
C
Forest
E
E
D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000378:metabolism; D000954:Antigens, Surface / Q000737:chemistry* / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000276:immunology*; D001428:Bacterial Vaccines; D015153:Blotting, Western; D015748:Borrelia burgdorferi Group / Q000737:chemistry*; D002460:Cell Line; D002847:Chromatography, Agarose; D003488:Cyanogen Bromide / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry*; D005260:Female; D005998:Glycine / Q000737:chemistry; D007477:Ions; D008074:Lipoproteins; D022123:Lyme Disease Vaccines / Q000737:chemistry* / Q000276:immunology*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000941:Antigens; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D000939:Epitopes; D007477:Ions; D008074:Lipoproteins; D022123:Lyme Disease Vaccines; C072045:OspA protein; D010455:Peptides; D011994:Recombinant Proteins; D003488:Cyanogen Bromide; D005998:Glycine