Autoepitope mapping of the centrosome autoantigen PCM-1 using scleroderma sera with anticentrosome autoantibodies.
We previously characterized a scleroderma serum (serum 1) containing autoantibodies against centrosome autoantigens that have been named PCM-1, PCM-2 and PCM-3. In this study, we analyzed another scleroderma serum (serum 2) reactive with centrosome autoantigens of identical molecular weights to those recognized by serum 1. To further analyze the autoepitope domains in PCM-1 recognized by the autoantibodies present in scleroderma sera, cDNAs encoding different portions of the PCM-1 autoantigen were expressed in bacteria as fusion proteins. The immunoreactivity of the fusion proteins to the scleroderma sera was assayed by immunoblot analysis. Two regions containing autoepitope domains reactive with both sera were identified in the PCM-1 molecule. One is between amino acids 312-706 of the PCM-1 autoantigen, and the other is localized between amino acids 1,433-1,787, indicating that the immune response is oligoclonal. The results are important to clarify the mechanism of induction of anticentrosome autoantibodies. The potential diagnostic and prognostic significance of the autoantibodies for subgroups of scleroderma is discussed.
Bao
L
L
Department of Structural and Cellular Biology, University of South Alabama, Mobile 36688, USA.
Zimmer
W E
WE
Balczon
R
R
D000818:Animals; D001323:Autoantibodies / Q000097:blood*; D001324:Autoantigens / Q000737:chemistry* / Q000276:immunology*; D016466:CHO Cells; D018797:Cell Cycle Proteins; D018385:Centrosome / Q000276:immunology*; D006224:Cricetinae; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D012595:Scleroderma, Systemic / Q000276:immunology*
D001323:Autoantibodies; D001324:Autoantigens; D018797:Cell Cycle Proteins; D000939:Epitopes; C086074:PCM1 protein, human