Mapping of SLE-specific Sm B cell epitopes using murine monoclonal antibodies.
In this study we have used a number of monoclonal antibodies with various anti-Sm specificities originating from MRL/lpr mice to map B cell epitopes of the Sm-B/B' and Sm-D1 proteins. Selection of Sm-B subfragments reactive with the Sm-B/B'-specific monoclonal antibody ANA125 from a DNaseI fragment expression library revealed that the epitope recognized by this monoclonal antibody is located between amino acids 146 and 158: GRGTVAAAAAAAT. The epitopes recognized by two distinct Sm-D1-specific monoclonal antibodies, 7.13 and ANA127, appeared to be located in the carboxy-terminal region of the protein as revealed by immunoprecipitation of in vitro translated deletion mutants of Sm-D1. These epitopes are probably identical and not simply composed of a GR repeat, which is a characteristic feature of this part of the protein. Immunoprecipitation of in vitro translated deletion mutants of both Sm-B and Sm-D1 was also employed to determine the sequence requirements for recognition by two monoclonal antibodies that are cross-reactive with several Sm proteins, Y12 and ANA128. The epitope recognized by these two monoclonal antibodies is probably also identical and composed by the juxtaposition of several regions in the folded protein. The low, but significant, level of immunoprecipitation of truncated versions of both Sm-B and Sm-D1, suggests that the Sm domain, which is shared by all Sm proteins, in particular the amino-terminal part of the Sm1 motif of Sm-B and Sm-D1, plays an important role in formation of the cross-reactive epitope and might contribute to cross-reactivity with other Sm proteins. The results of immunoprecipitation experiments with cellular extracts show that the epitopes recognized by all anti-Sm monoclonal antibodies used in this study are accessible in the assembled snRNPs.
Pruijn
G J
GJ
Department of Biochemistry, University of Nijmegen, The Netherlands.
Schoute
F
F
Thijssen
J P
JP
Smeenk
R J
RJ
van Venrooij
W J
WJ
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000918:Antibody Specificity; D001323:Autoantibodies / Q000737:chemistry; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology*; D003429:Cross Reactions; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000737:chemistry*; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology*; D051379:Mice; D019463:Mice, Inbred MRL lpr; D008969:Molecular Sequence Data; D019151:Peptide Library; D011487:Protein Conformation; D017411:Ribonucleoproteins, Small Nuclear / Q000737:chemistry / Q000276:immunology*; D055517:snRNP Core Proteins
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001324:Autoantigens; D018985:Epitopes, B-Lymphocyte; D019151:Peptide Library; D017411:Ribonucleoproteins, Small Nuclear; C526416:Snrpb protein, mouse; D055517:snRNP Core Proteins