这是一种快速的方法,可以在单个酵母细胞中区分发酵和呼吸代谢
Laura Luzia1, Julius Battjes1, Emile Zwering1
1Systems Biology Lab, A-LIFE, Institute of Molecular and Life Sciences (AIMMS), VU Amsterdam, 1081HZ Amsterdam, the Netherlands.
iScience
|January 18, 2024
概括
我们开发了iCRAFT,这是一种在单个酵母细胞中区分发酵和呼吸的方法. 这种单细胞方法在营养过渡过程中揭示了异质的代谢反应,进步了我们对酵母代谢的理解.
科学领域:
- 微生物学 微生物学
- 生物化学 生物化学
- 系统生物学 系统生物学
背景情况:
- 在葡萄糖耗尽的过程中, *Saccharomyces cerevisiae* 的新陈代谢从发酵转向呼吸.
- *目前的理解依赖于种群平均值,掩盖了单个细胞的变异性.
- * 异质的代谢反应对于理解酵母适应至关重要.
研究的目的:
- * 引入iCRAFT,一种用于酵母菌单细胞代谢分析的新方法.
- * 为了区分酵母细胞中的发酵和呼吸代谢.
- * 调查营养过渡期间的代谢异质性.
主要方法:
- *将基于ATP Förster共振能量转移 (FRET) 的生物传感器yAT1.03与Antimycin A.结合起来.
- *根据正常化的FRET比率变化,区分代谢状态.
- * 在*Saccharomyces cerevisiae**的*diauxic转移期间跟踪代谢变化.
主要成果:
- *iCRAFT在添加抗菌素A后成功区分了呼吸 (急剧降低FRET) 和呼吸发酵细胞 (没有反应).
- *单细胞分析显示,在葡萄糖耗尽后,整个人口的呼吸ATP产量逐渐增加.
- * 有证据表明,在单细胞水平上,呼吸能力逐渐增加.
结论:
- *iCRAFT是一种强大的工具,可以在单个酵母细胞中区分发酵和呼吸.
- * 该方法为研究代谢调节和适应提供了新的机会.
- *了解单细胞代谢是解读酵母复杂代谢调整的关键.
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