Pyruvate kinase

About: Pyruvate kinase is a research topic. Over the lifetime, 5683 publications have been published within this topic receiving 180020 citations. The topic is also known as: ATP:pyruvate 2-O-phosphotransferase & phosphoenolpyruvate kinase.

Mechanism of action of glucagon on hepatocyte phosphofructokinase activity.

Abstract: Addition of glucagon to isolated hepatocytes reduced the activity of 6-phosphofructokinase (ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) and pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40). Phosphorylation contributed to the inhibition of pyruvate kinase, but several lines of evidence indicated that this reaction was not responsible for the inhibition of phosphofructokinase. First, the increase in phosphorylation in intact cells induced by increasing the concentration of glucagon did not correlate well with the decrease in enzyme activity. Second, phosphorylation of phosphofructokinase induced by addition of cyclic AMP and Mg2+-ATP or by addition of Mg2+-ATP and the catalytic subunit of the cyclic AMP-dependent protein kinase to hepatocyte extracts had no effect on enzyme activity. Third, ammonium sulfate precipitation of the enzyme from extracts of cells incubated with glucagon abolished the hormone effect. The effect could be restored, however, by the addition of a phosphofructokinase-free extract from glucagon-treated cells to the ammonium sulfate-treated enzyme from either untreated or glucagon-treated cells. These results suggest that the inhibition of phosphofructokinase by glucagon is due to changes in the level of an allosteric effector(s).

Glycolysis and the regulation of cryoprotectant synthesis in liver of the freeze tolerant wood frog

Abstract: Wood frogs,Rana sylvatica, were sampled after freezing at −4°C (a short time course from 2 to 70 min after the appearance of the freezing exotherm) and thawing (20 h at 3°C after 70 min of freezing) and the regulation of liver glycolysis with respect to cryoprotectant glucose synthesis was examined. Within 5 min of the initiation of freezing, cryoprotectant concentrations in blood and liver had begun to increase. This was correlated with a rapid rise in the levels of hexose monophosphates in liver, including a 2.5 fold increase in glucose-6-P and 10 fold rise in fructose-6-P contents within the first 5 min post-exotherm. Contents of fructose-1,6-P2, fructose-2,6-P2, triose phosphates, P-enolpyruvate, and pyruvate did not significantly change over the course of freezing. Thawing sharply reduced the levels of hexose monophosphates in liver but raised P-enolpyruvate content by 2.3 fold. Changes in the contents of glycolytic intermediates over the freeze/thaw course are consistent with an inhibitory block of glycolysis at phosphofructokinase during freezing in order to facilitate a rapid glycogenolysis and production of cryoprotectant; during thawing, however, glycolysis appears to be inhibited at the level of pyruvate kinase. Possible regulatory control of cryoprotectant synthesis by covalent modification of liver glycolytic enzymes was examined. Glycogenolysis during freezing was facilitated by an increase in the percentage of glycogen phosphorylase in the activea (phosphorylated) form and also by an increase in the total amount (a+b) of enzyme expressed. For phosphofructokinase, kinetic changes as a result of freezing included a 40% reduction inK m for fructose-6-P, a 60% decrease inK a for fructose-2,6-P2, and a 2 fold increase in I50 for ATP. These changes imply a freezing-induced covalent modification of the enzyme but are not, apparently, the factors responsible for inhibition of glycolytic flux at the phosphofructokinase locus during glucose synthesis. Kinetic parameters of pyruvate kinase were not altered over the freeze/thaw course.

Role of AMP-activated protein kinase in the glycolysis of postmortem muscle

Abstract: AMP-activated protein kinase (AMPK) is a newly identified kinase controlling energy metabolism in vivo. The objective of this study was to show the role of AMPK in postmortem glycolysis. Rapid and excessive postmortem glycolysis is directly related to the incidence of PSE (pale, soft and exudative) meat in pork, chicken and turkey, while insufficient glycolysis leads to dark cutters in beef and lamb, which causes significant loss to the meat industry. A total of 24 two-month-old C57BL/6J mice were assigned to three treatments: (1) wild-type mice without pre-slaughter treatment; (2) wild-type mice with a 2 min swim before slaughter; and (3) wild-type mice intraperitoneally injected with AICAr (50 mg kg−1), a specific activator of AMPK, to stimulate the activity of AMPK. In addition, 16 two-month-old C57BL/6J mice with AMPK knockout were assigned to two treatments: (4) AMPK knockout mice without pre-slaughter treatment; and (5) AMPK knockout mice with a 2 min swim before slaughter. The longissimus dorsi muscle was sampled at 0, 1 and 24 h postmortem for pH and enzyme activity measurements. Results showed that AMPK activity had a major role in determining the ultimate muscle pH. Pre-slaughter stress induced by swimming significantly accelerated the glycogenolysis in postmortem muscle through activating glycogen phosphorylase. AMPK is important for maintaining the activity of glycogen phosphorylase and pyruvate kinase, and glycogenolysis/glycolysis in postmortem muscle. Thus, AMPK has an important role in the control of postmortem glycolysis and is crucial for a lower ultimate pH in postmortem muscle. However, the activation of AMPK cannot fully account for the initial rapid glycogenolysis/glycolysis induced by stress and another mechanism must exist for the accelerated glycolysis induced by pre-slaughter stress. Copyright © 2005 Society of Chemical Industry