Teusink B, Larsson C, Diderich J, Richard P, van Dam K, Gustafsson L, Westerhoff H V
E. C. Slater Institute, BioCentrum, University of Amsterdam, Plantage Muidergracht 12, NL-1018 TV Amsterdam, The Netherlands.
J Biol Chem. 1996 Oct 4;271(40):24442-8. doi: 10.1074/jbc.271.40.24442.
Microcalorimetry was adapted to the study of glycolytic oscillations in suspensions of intact yeast cells. A correction procedure was developed for the distortion of the amplitude and phase of the heat signal, caused by the slow response of the calorimeter. This made it possible to observe oscillations in the heat production rate with a period of less than 1 min, and a relative amplitude of 5-10%. By simultaneously measuring the heat flux and concentrations of glycolytic metabolites, and by comparing acetaldehyde-induced phase shifts of the heat flux oscillations with those of NADH oscillations, the heat flux was found to be 100 degrees out of phase with glucose 6-phosphate, 80 degrees out of phase with fructose 1, 6-bisphosphate, and in phase with NADH. The flux measurement made possible by microcalorimetry allowed the recognition of (i) changes in metabolic capacity that may affect glycolytic dynamics, (ii) implications of glucose carrier kinetics for glycolytic dynamics and (iii) the continued requirement for an acetaldehyde trapping agent for the oscillations.
微量量热法被应用于完整酵母细胞悬液中糖酵解振荡的研究。针对量热计的缓慢响应导致的热信号幅度和相位失真，开发了一种校正程序。这使得观察到产热速率的振荡成为可能，其周期小于1分钟，相对幅度为5 - 10%。通过同时测量热通量和糖酵解代谢物的浓度，并将乙醛诱导的热通量振荡的相移与NADH振荡的相移进行比较，发现热通量与6 - 磷酸葡萄糖相差100度，与1,6 - 二磷酸果糖相差80度，与NADH同相。微量量热法实现的通量测量使得能够识别：(i) 可能影响糖酵解动力学的代谢能力变化；(ii) 葡萄糖载体动力学对糖酵解动力学的影响；以及(iii) 振荡对乙醛捕获剂的持续需求。
