Molecular cloning of a major CENP-B epitope and its use for the detection of anticentromere autoantibodies.
An enzyme-linked immunosorbent assay (ELISA) has been developed for the detection of anticentromere autoantibodies in sera of patients with suspected or manifest rheumatic diseases. The antigen source used in this assay consists of the recombinant protein of glutathione S-transferase (GST) fused to the last 60 C-terminal amino acid residues of the major centromere protein CENP-B. Although this CENP-B segment is only a small part of the complete polypeptide, we show that it constitutes an important autoimmune antigenic domain which is recognized by all patient sera in which ACA can be detected using the immunoblotting technique with a HeLa S3 nuclear protein extract as antigen source.
Verheijen
R
R
Department of Biochemistry, University of Nijmegen, The Netherlands.
de Jong
B A
BA
Oberyé
E H
EH
van Venrooij
W J
WJ
D000595:Amino Acid Sequence; D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D001324:Autoantigens / Q000235:genetics* / Q000276:immunology; D002503:Centromere / Q000276:immunology*; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics* / Q000276:immunology; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012216:Rheumatic Diseases / Q000235:genetics / Q000276:immunology*
D001323:Autoantibodies; D001324:Autoantigens; C497926:CENPB protein, human; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D004247:DNA; D005982:Glutathione Transferase