Autoreactive epitopes within the human alpha-enolase and their recognition by sera from patients with endometriosis.
Patients with endometriosis significantly develop autoantibodies directed against endometrial proteins, which may be involved in the aetiology of this gynaecological disease. Based on standard Western blot analysis, a 48 kDa protein was localized in the soluble protein extract of endometrial adenocarcinoma cells using sera from patients with clinically staged endometriosis and identified as the glycolytic enzyme alpha-enolase. The corresponding cDNA coding for the human alpha-enolase was isolated from a human endometrial cDNA library and cloned into the vector pH6EX3, allowing the efficient expression of recombinant human alpha-enolase with an N-terminal histidine-hexapeptide as affinity ligand in Escherichia coli. The purified recombinant human alpha-enolase was evaluated as a specific antigenic tool for the diagnostic measurement of antiendometrial antibodies in sera from patients with endometriosis. With selected endometriosis sera, two linear autoreactive epitopes were localized within the recombinant human alpha-enolase using epitope mapping techniques, and they were characterized.
Walter
M
M
Department of Molecular Biology, ELIAS Entwicklungslabor, Freiburg, Germany.
Berg
H
H
Leidenberger
F A
FA
Schweppe
K W
KW
Northemann
W
W
D000230:Adenocarcinoma / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D000595:Amino Acid Sequence; D001324:Autoantigens / Q000235:genetics / Q000276:immunology*; D015551:Autoimmunity; D001483:Base Sequence; D015153:Blotting, Western; D018076:DNA, Complementary / Q000235:genetics; D016889:Endometrial Neoplasms / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D004715:Endometriosis / Q000097:blood / Q000276:immunology*; D018604:Epitope Mapping; D005260:Female; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D001324:Autoantigens; D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D010751:Phosphopyruvate Hydratase