Campbell-Burk S L, den Hollander J A, Alger J R, Shulman R G
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06511.
Biochemistry. 1987 Nov 17;26(23):7493-500. doi: 10.1021/bi00397a044.
31P NMR saturation-transfer techniques have been employed in glucose-grown derepressed yeast to determine unidirectional fluxes in the upper part of the Embden-Meyerhof-Parnas pathway. The experiments were performed during anaerobic and aerobic glycolysis by saturating the ATP gamma resonances and monitoring changes in the phosphomonoester signals from glucose 6-phosphate and fructose 1,6-bis-phosphate. These experiments were supplemented with 13C NMR measurements of glucose utilization rates and 13C NMR label distribution studies. Combined with data obtained previously from radioisotope measurements, these 31P and 13C NMR kinetic studies allowed estimation of the net glycolytic flow in addition to relative flows through phosphofructokinase (PFK) and Fru-1,6-P2ase during anaerobic and aerobic glycolysis. The 31P NMR saturation-transfer results are consistent with previous results obtained from measurements of metabolite levels, radioisotope data, and 13C NMR studies [den Hollander, J.A., Ugurbil, K., Brown, T.R., Bednar, M., Redfield, C., & Shulman, R.G. (1986a) Biochemistry 25, 203-211], providing additional support for in vivo measurement of the flows during glycolysis.
31P核磁共振饱和转移技术已用于葡萄糖培养的去阻遏酵母，以确定糖酵解途径上部的单向通量。实验在无氧和有氧糖酵解过程中进行，通过饱和ATP的γ共振并监测6-磷酸葡萄糖和1,6-二磷酸果糖的磷酸单酯信号变化。这些实验辅以葡萄糖利用率的13C核磁共振测量和13C核磁共振标记分布研究。结合先前从放射性同位素测量获得的数据，这些31P和13C核磁共振动力学研究除了能估计无氧和有氧糖酵解过程中通过磷酸果糖激酶（PFK）和果糖-1,6-二磷酸酶的相对通量外，还能估计糖酵解的净流量。31P核磁共振饱和转移结果与先前从代谢物水平测量、放射性同位素数据和13C核磁共振研究中获得的结果一致[登·霍兰德，J.A.，乌古尔比尔，K.，布朗，T.R.，贝德纳，M.，雷德菲尔德，C.，&舒尔曼，R.G.（1986a）《生物化学》25，203 - 211]，为糖酵解过程中通量的体内测量提供了额外支持。
