Solubilization and characteristics of the thyroid NADPH-dependent H2O2 generating system.
Solubilization of the thyroid particulate-associated NADPH-dependent H2O2 generating system has been tested with different detergents; (3-(3-cholamidopropyl)-dimethylammonio)1-propane sulfonate (CHAPS) was found to be the best of the six detergents tested. The ratio of H2O2 generation to NADPH oxidation was similar for CHAPS extract and native particulate material. CHAPS was also the only detergent able to preserve the Ca++-sensitivity of the NADPH oxidase. Solubilization of this enzyme allowed the determination of some of its characteristics: specificity for divalent cations, apparent Km for NADPH, optimum pH and sensitivity to SH- reagents.
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002413:Cations, Divalent; D003902:Detergents; D006861:Hydrogen Peroxide / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D019255:NADPH Oxidases; D012995:Solubility; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013961:Thyroid Gland / Q000201:enzymology*
D002413:Cations, Divalent; D003902:Detergents; D013439:Sulfhydryl Reagents; D006861:Hydrogen Peroxide; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases; D002118:Calcium