Improved assay procedures for thyroid peroxidase; application to normal and adenomatous human thyroid tissue.
We have developed assays for thyroid peroxidase in crude thyroid tissue preparations, in which a linear relationship between activity and amount of tissue could be demonstrated. Linear assays were developed based on the following peroxidase catalyzed reactions in the presence of H2O2:(1) oxidation of I- to I(-3), (2) oxidation of guaiacol, and (3) iodination of human goiter thyroglobulin. To attain satisfactory linearity we found it necessary to solubilize the enzyme beforehand. This was accomplished by a brief treatment of the particulate fraction with trypsin and deoxycholate, followed by centrifugation at 40 000 X g and dialysis. Not only did this treatment facilitate the development of linear assays, but it also resulted in a substantial increase in enzyme activity compared with that in the untreated particulate fraction. The use of a Polytron homogenizer for the initial disruption of the tissue also proved helpful in developing these assay procedures. The three different assays were used to measure peroxidase activities in human thyroid adenomas and in normal tissue derived from adenomatous glands. T he adenomas generally displayed a higher level of peroxidase activity than normal tissue. The greatest difference was observed with the iodination assay and the smallest difference with the guaiacol assay.
D000236:Adenoma / Q000201:enzymology*; D000818:Animals; D006139:Guaiacol; D006801:Humans; D007453:Iodide Peroxidase / Q000032:analysis*; D007455:Iodine; D008722:Methods; D010544:Peroxidases / Q000032:analysis*; D013552:Swine; D013961:Thyroid Gland / Q000201:enzymology*; D013964:Thyroid Neoplasms / Q000201:enzymology*
D006139:Guaiacol; D007455:Iodine; D010544:Peroxidases; D007453:Iodide Peroxidase