Reibstein D, den Hollander J A, Pilkis S J, Shulman R G
Biochemistry. 1986 Jan 14;25(1):219-27. doi: 10.1021/bi00349a031.
The kinetics of yeast phosphofructo-1-kinase has been studied in vitro. Effector concentrations (Fru-6-P, ATP, ADP, AMP, Pi, Fru-1,6-P2, and Fru-2,6-P2) and pH were adjusted so as to mimic intracellular concentrations in yeast. Under these conditions we were able to reproduce the measured in vivo rate of PFK. In addition, by reconstituting the intracellular conditions existing during aerobic and anaerobic glycolysis, we were able to reproduce in vitro the changes in the rate of PFK observed under these conditions. Without the addition of the newly discovered effector Fru-2,6-P2, in vitro rates of PFK are much lower than its in vivo rate. Changes in Fru-2,6-P2, Fru-1,6-P2, ATP, AMP, Pi, and pH in going from aerobic to anaerobic conditions all contributed somewhat to the change in the rate of PFK observed during the Pasteur effect, with no contribution coming from ADP. These studies show that the control of PFK under the condition of the Pasteur effect cannot be ascribed to changes in any one particular effector but rather to contributions from a variety of effectors. Also, the net change in the rate of PFK in the switch from anaerobic to aerobic glycolysis is small compared with the change in its dependence upon its substrate Fru-6-P, indicating a compensation mechanism.
已在体外研究了酵母磷酸果糖激酶-1的动力学。调节效应物浓度（果糖-6-磷酸、ATP、ADP、AMP、磷酸、果糖-1,6-二磷酸和果糖-2,6-二磷酸）和pH值，以模拟酵母细胞内的浓度。在这些条件下，我们能够重现所测得的磷酸果糖激酶的体内活性。此外，通过重建有氧和无氧糖酵解过程中存在的细胞内条件，我们能够在体外重现这些条件下观察到的磷酸果糖激酶活性的变化。如果不添加新发现的效应物果糖-2,6-二磷酸，磷酸果糖激酶的体外活性远低于其体内活性。从有氧条件转变为无氧条件时，果糖-2,6-二磷酸、果糖-1,6-二磷酸、ATP、AMP、磷酸和pH值的变化都对巴斯德效应期间观察到的磷酸果糖激酶活性的变化有一定贡献，而ADP没有贡献。这些研究表明，巴斯德效应条件下磷酸果糖激酶的调控不能归因于任何一种特定效应物的变化，而是多种效应物共同作用的结果。此外，从无氧糖酵解转变为有氧糖酵解时，磷酸果糖激酶活性的净变化与其对底物果糖-6-磷酸依赖性的变化相比很小，这表明存在一种补偿机制。
