The specificity of human autoantibodies that react with both cell nuclei and plasma membranes: the nuclear antigen is present on core mononucleosomes.
We have examined the nature of the nuclear antigen recognized by certain natural human antibodies that react specifically with both cell nuclei and plasma membranes from many species. Partial purification of these antibodies, called X-ANA, is achieved by binding to and rapid elution from the surface of viable human leukocytes. Chicken erythrocyte chromatin was solubilized by digestion with staphylococcal nuclease and fractionated into a 0.15 M NaCl soluble fraction that consisted of core mononucleosomes lacking H1/H5, and a 0.15 M NaCl insoluble fraction composed of polynucleosomes with H1/H5 present. No proteins other than histones were detected. Native and reconstituted mononucleosomes displaced IgG of the leukocyte eluates from nuclei of frozen mouse kidney sections and from the walls of plastic tubes coated with polynucleosomes. The reconstituted core mononucleosomes were 4- 10-fold less efficient inhibitors than native mononucleosomes. Trypsin digested mononucleosomes, free high m.w. DNA, and free histones displayed no or very weak inhibitory activity. The data indicate that X-ANA recognize a complex consisting of the core histones H2A, H2B, H3, H4, and DNA of 140 to 200 base pairs in length.
D000974:Antibodies, Antinuclear / Q000302:isolation & purification*; D000918:Antibody Specificity; D001323:Autoantibodies / Q000302:isolation & purification*; D002462:Cell Membrane / Q000276:immunology; D002467:Cell Nucleus / Q000276:immunology; D002843:Chromatin / Q000302:isolation & purification; D004247:DNA / Q000494:pharmacology; D000939:Epitopes; D006657:Histones / Q000494:pharmacology; D006801:Humans; D009707:Nucleosomes / Q000276:immunology* / Q000378:metabolism; D011863:Radioimmunoassay; D014357:Trypsin / Q000494:pharmacology
D000974:Antibodies, Antinuclear; D001323:Autoantibodies; D002843:Chromatin; D000939:Epitopes; D006657:Histones; D009707:Nucleosomes; D004247:DNA; D014357:Trypsin