Properties of detergent-dispersed iodothyronine 5- and 5'-deiodinase activities from rat liver.
In order to obtain more knowledge about the regulation and mechanism of thyroid hormone deiodination, some properties of detergent-solubilized iodothyronine deiodinase have been studied. Moreover, a starting point for its purification has been made. Several chromatography media were tested for their ability to purify the deiodinases. In some instances, a 4-fold purification was obtained. Treatment of cholate-solubilized microsomes with 35% ammonium sulphate resulted in quantitative precipitation of the deiodinase activities and concomitant removal of phospholipid. The pellet could be solubilized with 0.3% W-1 ether and the deiodinase in this ammonium sulphate extract exhibited approximately 10-fold higher apparent Km and Vmax values for its substrate compared with the cholate extract. Readdition of some phospholipids was shown to decrease enzyme activity. Isoelectric focusing of W-1 ether-solubilized microsomes resulted in a major activity peak around pH 6.4 and a minor peak at pH 5.2, while in the ammonium sulphate extract the deiodinase had an isoelectric point at pH 9.3. Refocusing of this activity peak yielded a preparation with a specific activity only 3-times higher than in the ammonium sulphate extract. However, after sodium dodecyl sulphate polyacrylamide gel electrophoresis only five bands could be detected. The elevated kinetic parameters as well as the higher isoelectric point of the deiodinase after ammonium sulphate treatment were caused by delipidation of the enzyme. Both the change in isoelectric point and the behaviour on several column materials were found to be similar for the 5- and 5'-deiodinase activities. These results suggest that a single enzyme is operative in the deiodination of iodothyronines in rat liver and that its activity may be regulated by phospholipids.
D000818:Animals; D003902:Detergents; D005026:Ethylene Glycols / Q000494:pharmacology; D007453:Iodide Peroxidase / Q000378:metabolism*; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology*; D010544:Peroxidases / Q000378:metabolism*; D010743:Phospholipids / Q000494:pharmacology; D051381:Rats; D012995:Solubility
D003902:Detergents; D005026:Ethylene Glycols; D010743:Phospholipids; D010544:Peroxidases; D007453:Iodide Peroxidase