Richard P, Teusink B, Westerhoff H V, van Dam K
E.C. Slater Institute, University of Amsterdam, The Netherlands.
FEBS Lett. 1993 Feb 22;318(1):80-2. doi: 10.1016/0014-5793(93)81332-t.
Under a limited set of hitherto incompletely defined conditions, inhibition of respiration has been shown to cause transient oscillations in NAD(P)H fluorescence of yeast cells. In this paper, we apply a new method [1992, Anal. Biochem. 204, 118-132] for extraction of intracellular metabolites. This method involves spraying the cells into -40 degrees C methanol; the neutral pH allows extraction of nearly all intracellular metabolites, including NADH. Close to the shift from glucose to ethanol as a growth substrate, the cells acquire a make-up amenable to sustained oscillations in intracellular concentrations of NADH and glycolytic intermediates such as glucose-6-phosphate. NADH was found to oscillate between 200 microM and 400 microM intracellular concentration. The cellular make-up determining the tendency to oscillate is 'remembered' by the cells after three hours of starvation.
在一组迄今尚未完全明确的有限条件下,已表明呼吸抑制会导致酵母细胞的NAD(P)H荧光出现短暂振荡。在本文中,我们应用一种新方法[1992年,《分析生物化学》204卷,第118 - 132页]来提取细胞内代谢物。该方法包括将细胞喷入-40摄氏度的甲醇中;中性pH值允许提取几乎所有细胞内代谢物,包括NADH。在从葡萄糖作为生长底物转变为乙醇作为生长底物的过程中,细胞获得了一种特性,使得细胞内NADH和糖酵解中间产物(如6 - 磷酸葡萄糖)的浓度能够持续振荡。发现细胞内NADH浓度在200微摩尔至400微摩尔之间振荡。在饥饿三小时后,细胞“记住”了决定振荡趋势的细胞组成。