Improved assay method for activity of thyroid peroxidase-catalysed coupling of iodotyrosine residues of thyroglobulin utilizing h.p.l.c. for analysis of iodothyronines.

Abstract

The coupling of iodotyrosine residues of thyroglobulin (Tg) catalysed by thyroid peroxidase (TPO) has scarcely been studied with respect to the TPO of abnormal human thyroid glands. The present paper proposes a rapid and convenient assay method applicable for determining the coupling activity of a sample of less than 500 mg from each patient's thyroid. The main characteristics of the method are as follows: (i) mitochondrial/microsomal fractions of thyroid glands were treated with sodium cholate plus trypsin, and the supernatants obtained by ultracentrifugation were directly used for the assay of coupling and peroxidase activity of TPO; (ii) the formation of iodotyrosine residues catalysed by TPO was performed by using chemically iodinated Graves'-disease Tg containing 41 iodine atoms per molecule and with a high iodotyrosine and a low iodothyronine content; (iii) newly synthesized iodothyronine residues (thyroxine, 3,5,3'-tri-iodothyronine, and 3,3',5'-tri-iodothyronine) were analysed by h.p.l.c. after hydrolysis of Tg with proteinases and extraction of iodothyronines with ethyl acetate.

• Authors: T Ohmori, O Tarutani, T Hosoya
• Journal: The Biochemical journal (Biochem J) Vol. 262 Issue 1 Pg. 209-14 (Aug 15 1989) ISSN: 0264-6021 [Print] England
• PMID: 2818564 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Thyroglobulin
• Iodide Peroxidase
• Monoiodotyrosine
• Thyroxine

Topics

• Chromatography, High Pressure Liquid
• Graves Disease (enzymology)
• Humans
• Iodide Peroxidase (metabolism)
• Methods
• Microsomes (enzymology)
• Monoiodotyrosine (metabolism)
• Thyroglobulin (metabolism)
• Thyroid Gland (enzymology)
• Thyroxine (analysis)