title: Anticentromere antibody specific to human cells directed against the CENP-B autoantigen.
abstract: We describe the generation of a new antipeptide antibody that binds to the centromeric region of human mitotic chromosomes. This antibody was raised against a synthetic peptide corresponding to the 481-493 amino acid sequence of the human CENP-B autoantigen. Immunofluorescence analysis revealed that this anti-CENP-B serum showed an identical pattern to the human CREST anticentromere autoantibody in both mitotic cells and interphase nuclei. Immunoblotting showed that this antibody reacts with the recombinant human CENP-B autoantigen, indicating that it is directed to the 80-kDa centromere polypeptide. We have used this serum to determine, by indirect immunofluorescence, whether CENP-B is conserved in different mammalian species. Surprisingly, the human antipeptide antibody does not react with the centromeric proteins of cultured mouse, hamster, or Indian muntjac cells. Because the CENP-B gene has been cloned in human and mouse, our results suggest that the CENP-B epitope used as an immunogen in this study is not ubiquitous in mammalian cells, and that we have most probably established a monospecific antibody to the human centromere.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad de Cádiz, Puerto Real, Spain.
authors.forename: L A
authors.initials: LA
authors.lastname: Bejarano
authors.forename: J
authors.initials: J
authors.lastname: Bolivar
authors.forename: M M
authors.initials: MM
authors.lastname: Valdivia
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001323:Autoantibodies / Q000235:genetics*; D001324:Autoantigens / Q000235:genetics*; D016466:CHO Cells; D002503:Centromere / Q000276:immunology*; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics*; D006224:Cricetinae; D004268:DNA-Binding Proteins; D003670:Deer; D000939:Epitopes / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012595:Scleroderma, Systemic / Q000235:genetics / Q000276:immunology*; D013045:Species Specificity
chemical_list: D001323:Autoantibodies; D001324:Autoantigens; C497926:CENPB protein, human; C497927:Cenpb protein, mouse; D051936:Centromere Protein B; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D000939:Epitopes; D011993:Recombinant Fusion Proteins