This Article Full Text (PDF) Submit a related Letter to the Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by FINKE, R. Articles by RAPOPORT, B. Search for Related Content PubMed PubMed Citation Articles by FINKE, R. Articles by RAPOPORT, B.

Evidence for the Highly Conformational Nature of the Epitope(s) on Human Thyroid Peroxidase that Are Recognized by Sera from Patients with Hashimoto's Thyroiditis^*

Thyroid Molecular Biology Laboratory, Veterans Administration Medical Center, and the University California San Francisco, California 94121

Address all correspondence and requests for reprints to: Basil Rapoport, M.B., Veterans Administration Medical Center (111T), 4150 Clement Street, San Francisco, California 94121.

To define the epitope(s) on human thyroid peroxidase (TPO) recognized by antibodies in the sera of patients with autoimmune thyroid disease, we constructed and screened a human TPO cDNA sublibrary containing 3.8 million random fragments of human TPO cDNA, each 200–500 basepairs in length. These fragments would code for TPO polypeptides of 66–166 amino acid residues. The validity of this approach was first tested with a murine monoclonal antibody against the denatured human thyroid microsomal antigen (TPO). Analysis of the nucleotide sequence of 14 clones selected from this library enabled molecular identification of the epitope recognized by this monoclonal antibody. In contrast to the data obtained with the monoclonal antibody, sera from patients with Hashimoto's thyroiditis containing polyclonal antimicrosomal/TPO antibodies did not recognize the TPO protein fragments generated by this library. These results differ from previous data obtained with recombinant human TPO fragments generated as bacterial fusion proteins. Our data suggest that, contrary to previous concepts, the natural B-cell epitope(s) on human TPO may be highly conformational (requiring a complex 3-dimensional structure) or may be discontinuous (formed by distant regions of the linear polypeptide chain being brought into apposition by protein folding).

^* This work was supported by the Deutscher Akademischer Austauschdienst (Grant 312 402 501/9), Bonn, West Germany; NIH Grants DK-36182 and DK-19289, and the Research Service of the V.A.

Received October 4, 1989.

This article has been cited by other articles:

				D. Bresson, M. Pugniere, F. Roquet, S. A. Rebuffat, B. N-Guyen, M. Cerutti, J. Guo, S. M. McLachlan, B. Rapoport, V. Estienne, et al. Directed Mutagenesis in Region 713-720 of Human Thyroperoxidase Assigns 713KFPED717 Residues as Being Involved in the B Domain of the Discontinuous Immunodominant Region Recognized by Human Autoantibodies J. Biol. Chem., September 10, 2004; 279(37): 39058 - 39067. [Abstract] [Full Text] [PDF]

				D. Bresson, M. Cerutti, G. Devauchelle, M. Pugniere, F. Roquet, C. Bes, C. Bossard, T. Chardes, and S. Peraldi-Roux Localization of the Discontinuous Immunodominant Region Recognized by Human Anti-thyroperoxidase Autoantibodies in Autoimmune Thyroid Diseases J. Biol. Chem., March 7, 2003; 278(11): 9560 - 9569. [Abstract] [Full Text] [PDF]

				J. Guo, X.-M. Yan, S. M. McLachlan, and B. Rapoport Search for the Autoantibody Immunodominant Region on Thyroid Peroxidase: Epitopic Footprinting with a Human Monoclonal Autoantibody Locates a Facet on the Native Antigen Containing a Highly Conformational Epitope J. Immunol., January 15, 2001; 166(2): 1327 - 1333. [Abstract] [Full Text] [PDF]

				G. D. Chazenbalk, Y. Wang, J. Guo, J. S. Hutchison, D. Segal, J. C. Jaume, S. M. McLachlan, and B. Rapoport A Mouse Monoclonal Antibody to a Thyrotropin Receptor Ectodomain Variant Provides Insight into the Exquisite Antigenic Conformational Requirement, Epitopes and in Vivo Concentration of Human Autoantibodies J. Clin. Endocrinol. Metab., February 1, 1999; 84(2): 702 - 710. [Abstract] [Full Text]

				B. Czarnocka, M. Janota-Bzowski, R. S. McIntosh, M. S. Asghar, P. F. Watson, E. H. Kemp, P. Carayon, and A. P. Weetman Immunoglobulin G{kappa} Antithyroid Peroxidase Antibodies in Hashimoto's Thyroiditis: Epitope-Mapping Analysis J. Clin. Endocrinol. Metab., August 1, 1997; 82(8): 2639 - 2644. [Abstract] [Full Text] [PDF]

				X. Zhang and P. Arvan Cell Type-dependent Differences in Thyroid Peroxidase Cell Surface Expression J. Biol. Chem., October 6, 2000; 275(41): 31946 - 31953. [Abstract] [Full Text] [PDF]