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강사욱,곽민규 한국미생물학회 2021 The journal of microbiology Vol.59 No.1
Glutathione reductase (Glr1) activity controls cellular glutathione and reactive oxygen species (ROS). We previously demonstrated two predominant methylglyoxal scavengers– NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and alcohol dehydrogenase 1 (Adh1)–in glutathione-depleted γ- glutamyl cysteinyl synthetase-disrupted Candida albicans. However, experimental evidence for Candida pathophysiology lacking the enzyme activities of Mgd1 and Adh1 on glutathione- dependent redox regulation remains unclear. Herein, we have aimed to demonstrate that glutathione-dependent enzyme activities coupled with cellular ROS changes is regulated by methylglyoxal accumulation in Δmgd1/Δadh1 double disruptants. Δmgd1/Δadh1 showed severe growth defects and G1-phase cell cycle arrest. The observed complementary and reciprocal methylglyoxal-oxidizing and methylglyoxalreducing activities between Δmgd1 and Δadh1 were not always exhibited in Δmgd1/Δadh1. Although intracellular accumulation of methylglyoxal and pyruvate was shown in all disruptants, to a greater or lesser degree, methylglyoxal was particularly accumulated in the Δmgd1/Δadh1 double disruptant. While cellular ROS significantly increased in Δmgd1 and Δadh1 as compared to the wild-type, Δmgd1/Δadh1 underwent a decrease in ROS in contrast to Δadh1. Despite the experimental findings underlining the importance of the undergoing unbalanced redox state of Δmgd1/Δadh1, glutathione- independent antioxidative enzyme activities did not change during proliferation and filamentation. Contrary to the significantly lowered glutathione content and Glr1 enzyme activity, the activity staining-based glutathione peroxidase activities concomitantly increased in this mutant. Additionally, the enhanced GLR1 transcript supported our results in Δmgd1/Δadh1, indicating that deficiencies of both Adh1 and Mgd1 activities stimulate specific glutathione-dependent enzyme activities. This suggests that glutathione-dependent redox regulation is evidently linked to C. albicans pathogenicity under the control of methylglyoxal-scavenging activities.
전자스핀공명스펙트럼에 의한 saccharomyces cerevisiae의 세포환 연구
임형순,강사욱 한국미생물학회 1988 미생물학회지 Vol.26 No.1
The intracellular free radicals produced at different stages of cell cycle of Saccharomyces cerevisiae ATCC 24858 were investigated by means of electron spin resonance(ESR) spectroscopy. The synchronized cells by repeated starvation and refeeding revealed different ESR spectral pattern compared to that of asynchronized cells. Each spectrum centered at g=2.005, which indicates free radicals. The relative spin concentration was maximat at the end of DNA increase. The variation of the relative spin concentration at each distinct stage of the cell cycle was evaluated in relation to ascorbate concentration, L-galactonolactone oxidase activity, and ascorbate oxidase activity. The highest activities of L-galactonolactone oxidase and ascorbate oxidase were detected just before and at the maximum of relative spin concentration, respectively. And ascorbate concentration fluctuated through each stage of cell cycle with the changes of relative spin concentration, L-galactonolactone oxidase activity, and ascorbate oxidase activity. Thus it is suggested that intracellular free radicals should be related to cell cycle, interacted with ascorbate, and may play an important role in the cell cycle of Saccharomyces cerevisiae.