Identification of an alpha-helical epitope region on the PM/Scl-100 autoantigen with structural homology to a region on the heterochromatin p25beta autoantigen using immobilized overlapping synthetic peptides.
The polymyositis-scleroderma overlap syndrome (PM/Scl) autoantigen is a nucleolar multiprotein particle, presumably participating in the maturation of 5.8S rRNAs. The major target antigens of this particle are two polypeptides with apparent molecular masses of 100 and 75 kDa. In this study we identified the major linear epitopes along the PM/Scl-100 protein sequence by probing overlapping oligopeptides with anti-PM/Scl autoantisera. A major epitope region was identified between amino acids 231 and 245 of the PM/Scl-100 polypeptide. Mutational analysis of the corresponding peptide LDVPPALADFIHQQR by glycine-walk followed by immunodetection of the resulting peptides indicated that amino acids 234, 237, 240, and 241 of the PM/Scl-100 autoantigen are essential for binding of the corresponding antibodies. These results allow us to propose a local alpha-helical secondary structure for the PM/Scl-100 major epitope region. A homology search with the peptide LDVPPALADFIHQQR against the Swiss-Model three-dimensional database reveals some topological homology of the PM/Scl-100 major epitope region with the heterochromatin modifier protein p25beta, a known autoantigen recognized by antibodies from a subset of scleroderma patients.
Blüthner
M
M
Institute of Molecular Genetics, University of Heidelberg, Germany. bluthner@sirius.mgen.uni-heidelberg.de
Mahler
M
M
Müller
D B
DB
Dünzl
H
H
Bautz
F A
FA
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000737:chemistry* / Q000276:immunology; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry*; D018604:Epitope Mapping; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry* / Q000276:immunology; D017433:Protein Structure, Secondary; D017386:Sequence Homology, Amino Acid
D001324:Autoantigens; D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D000090266:Chromobox Protein Homolog 5; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; C077044:EXOSC10 protein, human