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Purification and Crystallisation of the Autoantigen Thyroid Peroxidase from Human Graves’ Thyroid Tissue
Milligram quantities of the human membrane autoantigen thyroid peroxidase (TPO) have been purified to a high degree of homogeneity by a combination of detergent solubilisation, monoclonal antibody affinity, and ion exchange chromatography, from pooled Graves' disease thyroid glands. The purifie...
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Published in: | Biochemical and biophysical research communications 1997-05, Vol.234 (2), p.366-370 |
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container_title | Biochemical and biophysical research communications |
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creator | Gardas, A. Sohi, M.K. Sutton, B.J. McGregor, A.M. Banga, J.P. |
description | Milligram quantities of the human membrane autoantigen thyroid peroxidase (TPO) have been purified to a high degree of homogeneity by a combination of detergent solubilisation, monoclonal antibody affinity, and ion exchange chromatography, from pooled Graves' disease thyroid glands. The purified TPO of greater than 90% purity was enzymatically active as judged by its ability to oxidise guaiacol. Crystals of TPO have been grown from solutions of the protein solubilised in sodium deoxycholate, in the presence of ammonium sulphate. The crystals exhibited birefringence under polarised light, indicative of molecular order. Crystallisation of this large, membrane autoantigen represents the first step in delineating the complete three-dimensional structure of a human autoantigen involved in destructive thyroiditis. |
doi_str_mv | 10.1006/bbrc.1997.6600 |
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subjects | Autoantigens - isolation & purification Chromatography, Affinity Chromatography, Ion Exchange Crystallization Deoxycholic Acid Graves Disease - enzymology Graves Disease - immunology Guaiacol Humans Iodide Peroxidase - immunology Iodide Peroxidase - isolation & purification Solubility Substrate Specificity Thyroid Gland - enzymology Thyroid Gland - immunology |
title | Purification and Crystallisation of the Autoantigen Thyroid Peroxidase from Human Graves’ Thyroid Tissue |
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