Thyroperoxidase, but not the thyrotropin receptor, contains sequential epitopes recognized by autoantibodies in recombinant peptides expressed in the pUEX vector.
The sequential epitopes on the human thyroperoxidase (TPO) recognized by antibodies in the sera of patients with autoimmune thyroid disease were investigated using a recombinant DNA technique. Previous studies led to the isolation of two overlapping cDNA clones that encode polypeptides of TPO (85 residues, C2; 100 residues, C21) recognized by sera from several patients with autoimmune disease that contained antimicrosomal autoantibodies. In this report the vector pUEX1 was used to clone and express small random fragments of TPO cDNA in Escherichia coli as a beta-galactosidase fusion protein. Colonies were screened with a serum from a patient with Hashimoto's thyroiditis, and immunoreactive peptides were identified by sequencing the corresponding DNA inserts. Two linear epitopes of human TPO (amino acids 590-622 and 710-722) were recognized by the autoantibodies. This confirmed our previous results and provide a more precise localization of the antigenic determinants involved. The same approach has been applied in an attempt to identify the binding site(s) for autoantibodies on the human TSH receptor. In contrast to the data obtained with TPO, sera from patients with blocking (from idiopathic myxoedema) or stimulating (from Graves' disease) activity did not recognize the linear TSH receptor peptide fragments generated in our libraries.
Libert
F
F
Institut de Recherche Interdisciplinaire en Biologie Humaine et Nucleaire, Brussels, Belgium.
Ludgate
M
M
Dinsart
C
C
Vassart
G
G
D000595:Amino Acid Sequence; D001323:Autoantibodies / Q000276:immunology*; D004247:DNA / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors; D006111:Graves Disease / Q000097:blood / Q000235:genetics; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D009230:Myxedema / Q000097:blood / Q000235:genetics; D010957:Plasmids; D011989:Receptors, Thyrotropin / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D013967:Thyroiditis, Autoimmune / Q000097:blood / Q000235:genetics
D001323:Autoantibodies; D000939:Epitopes; D011989:Receptors, Thyrotropin; D011994:Recombinant Proteins; D004247:DNA; D007453:Iodide Peroxidase