Heterogeneous nuclear ribonucleoproteins C1/C2 identified as autoantigens by biochemical and mass spectrometric methods.
The antigenic specificity of an unusual antinuclear antibody pattern in three patient sera was identified after separating HeLa-cell nuclear extracts by two-dimensional (2D) gel electrophoresis and localizing the antigens by immunoblotting with patient serum. Protein spots were excised from the 2D gel and their contents were analyzed by matrix-assisted laser desorption-ionization (MALDI) or nanoelectrospray ionization time-of-flight (TOF) tandem mass spectrometry (MS) after in-gel digestion with trypsin. A database search identified the proteins as the C1 and C2 heterogeneous nuclear ribonucleoproteins. The clinical spectrum of patients with these autoantibodies includes arthritis, psoriasis, myositis, and scleroderma. None of 59 patients with rheumatoid arthritis, 19 with polymyositis, 33 with scleroderma, and 10 with psoriatic arthritis had similar antibodies. High-resolution protein-separation methods and mass-spectrometric peptide mapping in combination with database searches are powerful tools in the identification of novel autoantigen specificities.
Heegaard
N H
NH
Department of Autoimmunology, Statens Serum Institut, Copenhagen, Denmark. nhe@ssi.dk
Larsen
M R
MR
Muncrief
T
T
Wiik
A
A
Roepstorff
P
P
D000328:Adult; D000368:Aged; D000974:Antibodies, Antinuclear / Q000097:blood; D001324:Autoantigens / Q000276:immunology / Q000302:isolation & purification*; D002457:Cell Extracts / Q000032:analysis; D002478:Cells, Cultured / Q000276:immunology; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D008297:Male; D008875:Middle Aged; D008970:Molecular Weight; D012216:Rheumatic Diseases / Q000276:immunology*; D012261:Ribonucleoproteins / Q000276:immunology / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000974:Antibodies, Antinuclear; D001324:Autoantigens; C439788:C1 HNRNP; D002457:Cell Extracts; C503635:HNRNPC protein, human; D034482:Heterogeneous-Nuclear Ribonucleoprotein Group C; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D012261:Ribonucleoproteins