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随着年龄增长生物分子的氧化以及酿酒酵母不同生长阶段诱导的氧化应激。

Oxidation of biological molecules with age and induced oxidative stress in different growth phases of Saccharomyces cerevisiae.

作者信息

Mukherjee Madhumathan, Jana Chandan Kumar, Das Nilanjana

机构信息

Department of Biotechnology, Visva-Bharati University, Santiniketan, West Bengal, 731235, India.

St. Teresa School, Santiniketan, Dist. Birbhum, 731235, India.

出版信息

Antonie Van Leeuwenhoek. 2023 Apr;116(4):353-365. doi: 10.1007/s10482-022-01807-8. Epub 2023 Feb 7.

Abstract

One of the mechanistic approaches for explaining ageing is the oxidative stress theory of ageing. Saccharomyces cerevisiae has been used as a model to study ageing due to many factors. We have attempted to investigate if the differential ability to withstand oxidative stress can be correlated with their lifespans. In all the four strains studied (AP22, 699, 8C, and SP4), there was no age-associated increases in lipid peroxidation levels measured as thiobarbituric acid reactive substances (TBARS). Under induced oxidative stress conditions, there was an increased TBARS level in both the ages assessed with a quantum-fold increase in the stationary phase cells of AP22. In contrast, the late stationary phase cells of 8C exhibited the least susceptibility to induced oxidative stress. The level of TBARS in both exponential and late stationary phase cells of 699 was overall more than that in the other three strains. Protein carbonylation increased with age in 8C and 699. Induced stress increased carbonylation in the exponential cells of SP4 and 699 and the stationary phase cells of all four strains. Protein carbonylation data indicate that the AP22 cells exhibit decreased protein carbonylation vis-à-vis the other strains. Induced stress data showed that while the exponential cells of 699 are susceptible, the late stationary phase cells of 699 are most resistant. Western blotting analysis using anti-HNE antibodies showed two proteins of molecular mass ~ 56 and ~ 84 kDa that were selectively modified with age in all the strains. Under induced stress conditions, an additional protein of ~ 69 kDa was oxidized. Our investigation shows that the difference in lifespan between the four strains of S. cerevisiae may be regulated by oxidative stress. Knowledge of the identity of the oxidized proteins will significantly facilitate a better understanding of the effect of oxidative stress conditions on the cells of S. cerevisiae.

摘要

解释衰老的一种机制方法是衰老的氧化应激理论。由于多种因素,酿酒酵母已被用作研究衰老的模型。我们试图研究承受氧化应激的差异能力是否与其寿命相关。在所研究的所有四种菌株(AP22、699、8C和SP4)中,以硫代巴比妥酸反应性物质(TBARS)衡量的脂质过氧化水平没有随年龄增加。在诱导的氧化应激条件下,在评估的两个年龄段中,AP22的稳定期细胞中TBARS水平均呈数量级增加。相比之下,8C的稳定期末期细胞对诱导的氧化应激表现出最小的敏感性。699的指数期和稳定期末期细胞中的TBARS水平总体上高于其他三种菌株。8C和699中的蛋白质羰基化随年龄增加。诱导应激增加了SP4和699的指数期细胞以及所有四种菌株的稳定期细胞中的羰基化。蛋白质羰基化数据表明,与其他菌株相比,AP22细胞的蛋白质羰基化减少。诱导应激数据表明,虽然699的指数期细胞敏感,但699的稳定期末期细胞最具抗性。使用抗HNE抗体的蛋白质印迹分析表明,所有菌株中分子量约为5^6和~84 kDa的两种蛋白质随年龄有选择性修饰。在诱导应激条件下,一种约69 kDa的额外蛋白质被氧化。我们的研究表明,酿酒酵母的四种菌株之间寿命的差异可能受氧化应激调节。了解氧化蛋白质的身份将显著有助于更好地理解氧化应激条件对酿酒酵母细胞的影响。

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