Activation of a murine autoreactive B cell by immunization with human recombinant autoantigen La/SS-B: characterization of the autoepitope.
Immunization of Balb/c mice with a homogeneously purified recombinant human La/SS-B protein resulted in activation of an autoreactive B cell secreting a novel monoclonal anti-La antibody termed La4B6. La4B6 reacted with La protein from a variety of sources including human, bovine, rat and mouse. ATP blocked the binding of La4B6 to recombinant La protein. The human epitope was identified as consisting of the amino acid sequence SKGRRFKGKGKGN, which includes the proposed ATP-binding site of the La protein. In the human and bovine La protein, the epitope exists as a continuous amino acid sequence. In rat and mouse the epitope was found to consist of the amino acid sequence SKG interrupted by a species-specific insert of 16 amino acids, and followed by the second half of the epitope, the amino acid sequence RRFKGKGKGN. Our data suggest that in the case of the rat and mouse La proteins the two separated parts of the epitope are able to form a conformational epitope which looks similar to the continuous human epitope.
Tröster
H
H
Institut für Physiologische Chemie, Johannes-Gutenberg Universität, Mainz, Germany.
Bartsch
H
H
Klein
R
R
Metzger
T E
TE
Pollak
G
G
Semsei
I
I
Schwemmle
M
M
Pruijn
G J
GJ
van Venrooij
W J
WJ
Bachmann
M
M
D016475:3T3 Cells; D000255:Adenosine Triphosphate / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000276:immunology*; D015551:Autoimmunity / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002417:Cattle; D002460:Cell Line; D018604:Epitope Mapping; D017353:Gene Deletion; D006801:Humans; D007114:Immunization; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D016716:PC12 Cells; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012261:Ribonucleoproteins / Q000276:immunology*; D000097585:SS-B Antigen
D000255:Adenosine Triphosphate; D001324:Autoantigens; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D000097585:SS-B Antigen