Yamashoji S, Ikeda T, Yamashoji K
Institute of King Brewing, Kakogawa, Japan.
Biochim Biophys Acta. 1991 Aug 2;1059(1):99-105. doi: 10.1016/s0005-2728(05)80191-4.
Luminol chemiluminescence was observed by addition of menadione to yeast cell suspension and was amplified 1000-fold by further addition of Fe-complex. Catalase, superoxide dismutase and ceruloplasmin had inhibitory effects on luminol chemiluminescence, indicating the extracellular generation of active oxygens (H2O2 and O2-) and reduction of Fe-complex. The generation of H2O2 and reduction of Fe-complex were mainly dependent on the activity of NADH: menadione oxidoreductase in the plasma membrane and cytosol fractions. Both luminol chemiluminescence and H2O2 production were sensitive to the inhibitory effects of proton conductor, ionophorous antibiotics and ATPase inhibitor rather than the inhibitors of the mitochondria electron transport system. The incubation of glucose with yeast cells caused a parallel increase in luminol chemiluminescence, H2O2 production and intracellular NADH concentration. These facts suggest that menadione-catalyzed H2O2 production and chemiluminescence are used as the indicators of cell activity to keep the NADH concentration and NADH: menadione oxidoreductase activity which may be sensitive to the change in pH and ion concentrations.
向酵母细胞悬液中加入甲萘醌可观察到鲁米诺化学发光，进一步加入铁络合物后其强度放大了1000倍。过氧化氢酶、超氧化物歧化酶和铜蓝蛋白对鲁米诺化学发光有抑制作用，表明细胞外有活性氧（H2O2和O2-）生成以及铁络合物被还原。H2O2的生成和铁络合物的还原主要依赖于质膜和胞质溶胶部分中NADH：甲萘醌氧化还原酶的活性。鲁米诺化学发光和H2O2的产生对质子导体、离子载体抗生素和ATP酶抑制剂的抑制作用敏感，而对线粒体电子传递系统抑制剂不敏感。酵母细胞与葡萄糖一起孵育会使鲁米诺化学发光、H2O2产生和细胞内NADH浓度同时增加。这些事实表明，甲萘醌催化的H2O2产生和化学发光可作为细胞活性的指标，以维持NADH浓度和NADH：甲萘醌氧化还原酶活性，而这可能对pH值和离子浓度的变化敏感。
