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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Bibliographic Details
Published in:Biochemical and biophysical research communications 1997-05, Vol.234 (2), p.366-370
Main Authors: Gardas, A., Sohi, M.K., Sutton, B.J., McGregor, A.M., Banga, J.P.
Format: Article
Language:English
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
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Description
Summary: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.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1997.6600