Activating effect of adenosine on rat erythrocyte glycolysis.

1. Adenosine increases the adenine nucleotide pool in rat erythrocytes. Hence, we tested the effect of the nucleoside on the glycolytic pathway in red blood cells. 2. A 2.5-fold increase in the level of fructose-1,6-bisphosphate and a 34% augmentation in lactate pool were observed in rat erythrocytes, 30 min after adenosine treatment. 3. Under conditions preventing adenosine metabolism, 1 microM nucleoside addition to isolated erythrocytes induced an 89% increase in lactate production and an increase in glucose consumption. 4. Activation of red cell phosphofructokinase (PFK) is produced by addition of microM concentrations of adenosine. Our data suggest a role for adenosine in the glycolysis flux regulation through PFK activation.     

Thermodynamics of red cell glycolysis.

1. Changes of Gibbs energy, enthalpy and entropy in red cells during steady-state glycolysis were discussed. 2. The heat production of red cells at various metabolic conditions was measured on a flow microcalorimeter with simultaneous analyses of lactate and other metabolites. The results were discussed in relation to enthalpy changes of the different metabolic steps in the glycolytic pathway.     

Reversal of glycolysis in yeast.

When a buffered, aerobic suspension of ethanol-grown cells of Saccharomyces cerevisiae is treated with ethanol, a rapid flux of metabolism is observed from endogenous phosphoenolpyruvate to hexose monophosphates. Intracellular concentrations of phosphoenolpyruvate, 2-phosphoglycerate, and 3-phosphoglycerate record a monotonic drop, while those of triose phosphates and fructose 1,6-diphosphate fall after an early rise; fructose 6-phosphate, mannose 6-phosphate, and glucose 6-phosphate levels rise to a plateau. Prior growth on glucose extinguishes fructose 1,6-diphosphatase activity and completely arrests the rise of the hexose monophosphates. By using mutants blocked at a number of glycolytic steps it has been concluded that the metabolic flow takes place along the Embden-Meyerhof pathway in the reverse direction bypassing pyruvate kinase and fructose 6-phosphate kinase. Ethanol acts as a trigger by supplying NADH at the glyceraldehyde 3-phosphate dehydrogenase step. The rate of the reversal in the span phosphoenolpyruvate to fructose 1,6-diphosphate approaches 40 μ mol of 3-carbon units per minute per gram of wet cells. The in vivo activity of fructose 1,6-diphosphatase is nearly a quarter of this rate.     

Post-mortem glycolysis in skeletal muscle. The extent of glycolysis in diluted preparation of mammalian muscle.

1. Sheep-, rabbit- and ox-muscle minces prepared soon after slaughter were diluted with 1 vol. of 0.16 m-potassium chloride in the absence (potassium chloride mince) and presence of added cofactor or glycolysable substrate, and the effects on the ultimate pH were examined. 2. Changes in the concentrations of glycogen and lactate and the concentrations of some phosphorus-containing fractions were determined in ox-muscle preparations. 3. Glycolysis ceased at appreciably higher pH in the potassium chloride mince than in undiluted mince. The inclusion of glycogen, ATP, ADP, NAD or magnesium chloride in the diluent had little effect on the ultimate pH of the diluted mince. 4. Lactic acid production continued at lower pH values in diluted mince containing added glucose 1-phosphate, fructose 1,6-diphosphate or glucose plus hexokinase than in potassium chloride mince. 5. The evidence points to failure of the phosphorylase step being responsible for the dilution effect.     

Inhibitory effect of D-glucosamine on glycolysis in bovine retina.

1. The effects of glucosamine concentration on the size of the lactate pool, on the levels of ATP, ADP, AMP and on the radioactivity incorporation from [1-14-C] glucosamine into lactate, N-acetylglucosamine and glucosamine-6-P were studied using whole bovine retinas. 2. The radioactive lactate, evaluated in relation to glucosamine molarity, after a modest initial increase, diminishes significantly. On the contrary the N-acetyl [1-14-C] glucosamine, the [1-14-C] glucosamine-6-P and, consequently, also the [1-14-C] glucosamine-6-P/[-14-C] lactate ratio increase with glucosamine molarity. 3. The retinal content of ATP shows a modest increment after incubation with low concentrations of D-glucosamine (0.5--2.0 mM) and a remarkable fall at higher concentrations. 4. Using retinal homogenates D-glucosamine clearly lowers the lactate production from glucose, glucose-6-P and fructose-1, 6-P2. 5. D-Glucosamine acts as an inhibitor of retinal glyceraldehyde-3-P dehydrogenase and lactate dehydrogenase by decreasing the initial velocity of these reactions. 6. It is concluded that D-glucosamine causes a reduction in the lactate production, by inhibiting two enzymes of the glycolytic pathway: glyceraldehyde-3-P dehydrogenase and lactate dehydrogenase. The fall in the adenine nucleotides content is a consequence of a dephosphorylation of ATP for the phosphorylation of glucosamine without concomitant resynthesis of ATP "via glycolysis".