Mapping of B cell epitopes on small nuclear ribonucleoproteins that react with human autoantibodies as well as with experimentally-induced mouse monoclonal antibodies.
Small nuclear ribonucleoprotein (snRNP) particles are a class of RNA-containing particles in the nucleus of eukaryotic cells. They consist of uridylate-rich small nuclear RNA complexed with several proteins. snRNP particles U1, U2, U4/U6, and U5 all contain a common protein core consisting of proteins B'/B, D, D', E, F, and G. In addition to this core, U1 snRNP particles contain proteins 70K, A, and C, whereas U2 snRNP particles contain proteins A' and B". Almost any of the small nuclear RNA-associated polypeptides is targeted by autoantibodies in the sera from patients with SLE or related connective tissue diseases. We immunized a genetically non-autoimmune mouse with recombinant human B" protein and obtained three mAb reactive with native U2 snRNP particles. Two of these mAb particles cross-reacted with U1 snRNP, 9A9 and 11A1, via epitopes present on the U2 snRNP B" protein as well as on the U1 snRNP-specific A protein. A third mAb 4g3, reacted exclusively with U2 snRNP via a unique epitope on protein B". Two epitopes mapped at the carboxy-terminal region of the B" protein, whereas binding of the third mAb involved both amino- and carboxy-terminal amino acids of the B" protein. Epitope mapping, employing a DNAse I fragment library of the B" cDNA, revealed that the three mAb-reactive sites were discontinuous. Autoantibodies in sera from patients with SLE and other connective tissue diseases competed for binding with the mAb, implying that the mAb define a major autoantibody-reactive region on protein B".
Habets
W J
WJ
Department of Biochemistry, University of Nijmegen, The Netherlands.
Hoet
M H
MH
De Jong
B A
BA
Van der Kemp
A
A
Van Venrooij
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D000944:Antigens, Differentiation, B-Lymphocyte / Q000032:analysis* / Q000276:immunology; D001323:Autoantibodies / Q000276:immunology*; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D003240:Connective Tissue Diseases / Q000276:immunology; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D012261:Ribonucleoproteins / Q000032:analysis* / Q000276:immunology; D017411:Ribonucleoproteins, Small Nuclear
D000911:Antibodies, Monoclonal; D000944:Antigens, Differentiation, B-Lymphocyte; D001323:Autoantibodies; D000939:Epitopes; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear