Molecular characterization of an autoantigen of PM-Scl in the polymyositis/scleroderma overlap syndrome: a unique and complete human cDNA encoding an apparent 75-kD acidic protein of the nucleolar complex.
About 50% of patients with the polymyositis/scleroderma (PM-Scl) overlap syndrome are reported to have autoantibodies to a nuclear/nucleolar particle termed PM-Scl. The particle is composed of several polypeptides of which two have been identified as autoantigens. In this report, human cDNA clone coding for the entire 75-kD autoantigen of the PM-Scl particle (PM-Scl 75) was isolated from a MOLT-4 lambda gt-11 library. The deduced amino acid sequence of the cDNA clone represented a protein of 355 amino acids and 39.2 kD; the in vitro translation product of this cDNA migrated in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at approximately 70 kD. The aberrant migration of the polypeptide in SDS-PAGE was shown to be related to the COOH half that was rich in acidic residues. Authenticity of the cDNA coding for PM-Scl 75 was shown by immunoreactivity of PM-Scl sera with in vitro translation products and recombinant fusion proteins encoded by the cDNA. In addition, rabbit antibodies raised to recombinant fusion protein reacted in immunofluorescence, immunoblotting, and immunoprecipitation with the characteristic features displayed by human anti-PM-Scl sera.
Alderuccio
F
F
Department of Molecular and Experimental Medicine, Scripps Clinic and Research Foundation, La Jolla, California 92037.
Chan
E K
EK
Tan
E M
EM
D000595:Amino Acid Sequence; D001324:Autoantigens / Q000235:genetics*; D001327:Autoimmune Diseases / Q000276:immunology*; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002466:Cell Nucleolus / Q000276:immunology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009220:Myositis / Q000276:immunology*; D009841:Oligonucleotides / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000276:immunology; D015183:Restriction Mapping; D012595:Scleroderma, Systemic / Q000276:immunology*; D013577:Syndrome; D014158:Transcription, Genetic
D001324:Autoantigens; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D004247:DNA