Phosphorus-31 nuclear magnetic resonance studies of the effect of oxygen upon glycolysis in yeast

Hollander, Jan A. den; Uğurbil, Kâmil; Tr, Brown; Shulman, Robert G.

doi:10.1021/bi00523a034

Cited by 207 publications

(109 citation statements)

References 35 publications

(29 reference statements)

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“…When glucose is metabolized aerobically, both the ATP turnover and the phosphate potential increased (see Fig. 6 and den Hollander et al, 1981Hollander et al, , 1986aCampbell-Burk et al, 1987). In this latter case, ethanol addition stimulated ATP turnover and raised the phosphate potential (Fig.…”

Section: Metabolic Conditionmentioning

confidence: 83%

Interactions between glucose metabolism and oxidative phosphorylations on respiratory‐competent Saccharomyces cerevisiae cells

Beauvoit

Rigoulet

Bunoust

et al. 1993

European Journal of Biochemistry

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The purpose of this work was to analyze the interactions between oxidative phosphorylations and glucose metabolism on yeast cells aerobically grown on lactate as carbon source and incubated in a resting cell medium. On such respiratory-competent yeast cells, four different metabolic steady states have particularly been studied: (a) glucose feeding under anaerobiosis, (b) ethanol supply under aerobiosis, (c) glucose supply under aerobiosis and (d) glucose plus ethanol under aerobiosis. For each condition, we measured: (a) the cellular ATP/ADP ratio and NADH content sustained under these conditions, (b) the glucose consumption rate (glucose conditions) and the respiratory rate (aerobic conditions).Under aerobic conditions, when ethanol is used as substrate, the ATP/ADP ratio and NADH level are very high as compared with glucose feeding. However, the rate of oxygen consumption is similar under both conditions. The main observation is a large increase in the respiratory rate when both glucose and ethanol are added. This increase corresponds to an ATP/ADP ratio and a NADH level lower than those observed with ethanol but higher than those with glucose. Therefore the response of the respiratory rate to the ATP/ADP ratio depends on the redox potential. We studied the way in which the ATP-consuming activity was increased under glucose + ethanol conditions. By NMR experiments, it appears that neither the futile cycle at the level of the phosphofructo-lkinase/fructo-l,6-bisphosphatase couple nor the synthesis of carbohydrate stores could account for the increase in oxidative phosphorylation. However, it is shown that, in the presence of glucose + ethanol, ATP consumption is strongly stimulated. It is hypothesized that this consumption is essentially due to the combination of the well-known plasma membrane proton-ATPase activation by glucose and the high phosphate potential due to oxidative ethanol metabolism. While it is well documented that oxidative phosphorylations inhibit the glycolytic flux, i.e. the Pasteur effect, we clearly show in this work that the glycolytic pathway limits the ability of mitochondria to maintain a cellular phosphate potential.Studies on isolated mitochondria have demonstrated that the ATP synthesis flux coupled to respiratory rate is controlled by the intramitochondrial phosphate potential value according to both the presence and the nature of the intra-or extramatricial ADP-regenerating system (Kuster et al

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Section: Metabolic Conditionmentioning

confidence: 83%

Interactions between glucose metabolism and oxidative phosphorylations on respiratory‐competent Saccharomyces cerevisiae cells

Beauvoit

Rigoulet

Bunoust

et al. 1993

European Journal of Biochemistry

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“…Furthermore, the in vivo transport of inorganic phosphate across the vacuolar membrane during aerobic glycolysis in Candida utilis will be discussed. So far, most 3~p NMR studies in yeasts have concentrated on S. cerevisiae (Navon et al 1979;Den Hollander et al 1981 ;Gillies et al 1981 ;Jacobson and Cohen 1981). Here we compare this organism with C. utilis and Z. bailii.…”

Section: Offprint Requests To : K Nicolaymentioning

confidence: 96%

Phosphorus-31 nuclear magnetic resonance studies of intracellular pH, phosphate compartmentation and phosphate transport in yeasts

et al. 1982

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Abstract. 31p NMR spectra were obtained from suspensions of Candida utilis, Saccharomyces cerevisiae and Zygosaccharomyces bailii grown aerobically on glucose. Direct introduction of substrate into the cell suspension, without interruption of the measurements, revealed rapid changes in pH upon addition of the energy source. All 3 ~p NMR spectra of the yeasts studied indicated the presence of two major intracellular inorganic phosphate pools at different pH environments. The pool at the higher pH was assigned to cytoplasmic phosphate from its response to glucose addition and iodoacetate inhibition of glycolysis. After addition of substrate the pH in the compartment containing the second phosphate pool decreased. A parallel response was observed for a significant fraction of the terminal and penultimate phosphates of the polyphosphate observed by 3 ~p NMR. This suggested that the inorganic phosphate fraction at the lower pH and the polyphosphates originated from the same intracellular compartment, most probably the vacuole. In this vacuolar compartment, pH is sensitive to metabolic conditions. In the presence of energy source a pH gradient as large as 0.8 to 1.5 units could be generated across the vacuolar membrane. Under certain conditions net transport of inorganic phosphate across the vacuolar membrane was observed during glycolysis: to the cytoplasm when the cytoplasmic phosphate concentration had become very low due to sugar phosphorylation, and into the vacuole when the former concentration had become high again after glucose exhaustion.

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“…Direct evidence for changes of the cytosolic adenylate system as a function of p02 is not available; changes of the total cellular adenine nucleotides were observed only below 30 pM oxygen (A 20 mmHg = 2.7 kPa or 3 % ) 116,401 and under anoxic conditions [41]. Furthermore, the redox status of the cell, which might be reflected by the NADf/NADH, the NADP+/NADPH [I61 and the disulfide/thiol ratio , and the intracellular pH [48] might be involved in the 02-dependent regulation of metabolism. The NAD'/NADH ratio, as measured with the cellular pyruvate/lactate ratio, remained constant down to 30 pM ( g 20 mmHg = 2.7 kPa or 3 x) and then decreased [16].…”

Section: Mechanism Of the 02-dependent Modulation Of Carbohydrate Metmentioning

confidence: 99%

Short‐term modulation of glycogen metabolism, glycolysis and gluconeogenesis by physiological oxygen concentrations in hepatocyte cultures

Wölfle

Schmidt

Jungermann

1983

European Journal of Biochemistry

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The influence of different oxygen concentrations (0 to 20 x, v/v) on the main pathways of carbohydrate metabolism was studied in rat hepatocyte cultures. Cells resembling the periportal or the perivenous cell type were obtained after 48 h culture under different hormonal conditions; they are referred to as 'periportal' or 'perivenous', respectively. Using radiochemical techniques the metabolic rates of the two cell types were measured between 48 h and 50 h under different oxygen tensions. Standard physiological substrates of 5 mM glucose, 2 mM lactate and endogenous glycogen were used.1. In cells incubated under 4 % 02, mimicking hepatovenous oxygen levels, compared to cells assayed under 13 % O,, mimicking arterial levels, glycogen degradation to free glucose and to lactate was faster while glycogen synthesis was slower; glycolysis, from glucose to lactate, was faster and gluconeogenesis, from lactate to glucose, was only slightly slower. 2. Under anoxic conditions glycogen breakdown was maximal and glycogen synthesis minimal; gluconeogenesis was also minimal, but glycolysis was not maximal, it reached its peak rate at 4 % 0,.3. COz-formation increased up to 6 4. Net glycogen metabolism: in 'perivenous' cells net glycogen synthesis was observed above, net glycogen degradation below 4 % 0 2 . In 'periportal' cells, which had a very low glycogen content, net glycogen metabolism was very small.5. Net glucose metabolism: in 'perivenous' cells net glucose formation was only seen under anoxic conditions. Net glucose utilization was observed at about the samc rate under all physiological 0, tensions. In 'periportal' cells net glucose formation increased clearly up to 6 6. Net lactate metabolism: in 'perivenous' cells net lactate formation occurred below, and net lactate utilization above, 6 % 0 2 . In 'periportal' cells a pronounced net lactate utilization was observed under all physiological 0 2 tensions.7. Net flow between glucose-6-phosphate and pyruvate was observed in the glycolytic direction in 'perivenous' and in the gluconeogenic direction in 'periportal' cells except under anoxic conditions. When O2 tensions were lowered, the percentage of 'futile cycling' was decreased in the 'perivenous', glycolytic hepatocytes; conversely, it was increased in the 'periportal', gluconeogenic hepatocytes.It is concluded that physiological oxygen concentrations modulate hepatic carbohydrate metabolism and that they contribute further to the proposed metabolic differences betwccn periportal and perivenous cells in vivo.and then stayed essentially constant at higher 0 2 tensions 0, and then remained almost constant. Enzymes. Fructose-1,6-bisphosphatase (EC 3.1 .I .11); glucokinase (EC 2.7.1.2); glucose-6-phosphalase (EC 3.1.3.9); phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32); pyruvatc kinase (EC 2.7.1.40); tyrosine aminotransferase (EC 2.6.1.5).4.7 kPa = 43 ~ 50 pM = 4-5 %) [I], which is less than that of large veins, 40 mmHg (= 5.3 kPa = 57 pM = 6 %) [2], Recently periportal to perivenous oxygen gradients have been obs...

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Phosphorus-31 nuclear magnetic resonance studies of the effect of oxygen upon glycolysis in yeast

Cited by 207 publications

References 35 publications

Interactions between glucose metabolism and oxidative phosphorylations on respiratory‐competent Saccharomyces cerevisiae cells

Interactions between glucose metabolism and oxidative phosphorylations on respiratory‐competent Saccharomyces cerevisiae cells

Phosphorus-31 nuclear magnetic resonance studies of intracellular pH, phosphate compartmentation and phosphate transport in yeasts

Short‐term modulation of glycogen metabolism, glycolysis and gluconeogenesis by physiological oxygen concentrations in hepatocyte cultures

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