Identification of protein components reactive with anti-PM/Scl autoantibodies.
The PM/Scl antigen from mammalian cells has been characterized as a nucleolar and nucleoplasmic molecular complex containing at least 16 polypeptides ranging in molecular weight from 110 to 20 kD. Of these polypeptides, we have found those of 68, 39 and 20 kD to be in a phosphorilated form. Whereas the entire complex was precipitated by all the anti-PM/Scl sera tested, in immunoblots the antibodies specifically recognized determinants on the 110-kD protein. This protein was immunoprecipitated more preferentially from nucleoli extracts than from total cell extracts. Moreover, this protein disappeared from the immunoprecipitates when treated with DNAse. Likewise, the immunoblot reaction of the specific antibodies with the 110-kD protein was abolished by treatment of the extracts with DNAse and trypsin, and was resistant when extracts were treated with RNAse. Affinity-purified antibodies from this protein selectively stained the nucleoli and the nucleoplasm of the mammalian cells. Moreover, when the cultured cells used in immunofluorescence were treated with DNAse, the affinity purified antibodies from the 110-kD protein gave negative fluorescence. However, when whole anti-PM/Scl sera were used, a nucleolar and nucleoplasmic staining was found. We conclude that the 110-kD protein has at least one of the autoimmunogenic epitopes of the PM/Scl antigen, recognized by all anti-PM/Scl sera tested. Other epitopes differing in their DNAse sensitivity may also be present in the PM/Scl antigen.
Department of Immunology, Santa Cruz y San Pablo Hospital, Barcelona, Spain.
C
C
Gelpi
A
A
Algueró
M
M
Angeles Martinez
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S
Vidal
C
C
Juarez
J L
JL
Rodriguez-Sanchez
D000918:Antibody Specificity; D001172:Arthritis, Rheumatoid / Q000276:immunology; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology*; D015153:Blotting, Western; D002466:Cell Nucleolus / Q000276:immunology*; D002843:Chromatin / Q000276:immunology; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; D005455:Fluorescent Antibody Technique; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000276:immunology; D008970:Molecular Weight; D010750:Phosphoproteins / Q000276:immunology; D011233:Precipitin Tests; D012216:Rheumatic Diseases / Q000276:immunology*; D012595:Scleroderma, Systemic / Q000276:immunology
D001323:Autoantibodies; D001324:Autoantigens; D002843:Chromatin; D010750:Phosphoproteins; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; C077044:EXOSC10 protein, human