甲状腺中依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的过氧化氢生成系统的增溶作用及特性

Solubilization and characteristics of the thyroid NADPH-dependent H2O2 generating system.

作者信息

Dupuy C, Virion A, Hammou N A, Kaniewski J, Dème D, Pommier J

出版信息

Biochem Biophys Res Commun. 1986 Dec 15;141(2):839-46. doi: 10.1016/s0006-291x(86)80249-2.

Abstract

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.

摘要

已用不同的去污剂测试了甲状腺微粒体相关的依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的过氧化氢生成系统的增溶作用；在测试的六种去污剂中，发现3-(3-胆酰胺丙基)-二甲基铵基-1-丙烷磺酸盐（CHAPS）效果最佳。CHAPS提取物和天然微粒体物质的过氧化氢生成与NADPH氧化的比率相似。CHAPS也是唯一能够保留NADPH氧化酶对钙离子敏感性的去污剂。该酶的增溶使得能够确定其一些特性：对二价阳离子的特异性、对NADPH的表观米氏常数（Km）、最适pH以及对巯基试剂的敏感性。