• BMB Reports
• /
• v.56 no.11
• /
• pp.618-623
• /
• 2023

Most cancer cells utilize glucose at a high rate to produce energy and precursors for the biosynthesis of macromolecules such as lipids, proteins, and nucleic acids. This phenomenon is called the Warburg effect or aerobic glycolysis- this distinct characteristic is an attractive target for developing anticancer drugs. Here, we found that Phosphofructokinase-1 (PFK-1) is a substrate of the Protein Phosphatase 4 catalytic subunit (PP4C)/PP4 regulatory subunit 1 (PP4R1) complex by using immunoprecipitation and in vitro assay. While manipulation of PP4C/PP4R1 does not have a critical impact on PFK-1 expression, the absence of the PP4C/PP4R1 complex increases PFK-1 activity. Although PP4C depletion or overexpression does not cause a dramatic change in the overall glycolytic rate, PP4R1 depletion induces a considerable increase in both basal and compensatory glycolytic rates, as well as the oxygen consumption rate, indicating oxidative phosphorylation. Collectively, the PP4C/PP4R1 complex regulates PFK-1 activity by reversing its phosphorylation and is a promising candidate for treating glycolytic disorders and cancers. Targeting PP4R1 could be a more efficient and safer strategy to avoid pleiotropic effects than targeting PP4C directly.

• Journal of Microbiology and Biotechnology
• /
• v.14 no.1
• /
• pp.56-61
• /
• 2004

A plasmid pSDK-l containing the Escherichia coli phosphofructokinase-l gene (pfkA) was constructed, and transferred into extremely acidophilic Acidithiobacillus thiooxidans Tt-7 by conjugation with the aid of plasmid RP4 at a frequency of $10^{-5}$ per recipient. This plasmid was stable in A. thiooxidans. The pfkA gene from E. coli could be expressed in this obligately autotrophic bacterium, but the enzyme activity (21.6 U/g protein) was lower than that in E. coli (K12: 85.9 Dig protein; DF1010 carrying plasmid pSDK-l: 96.6 U/g protein). In the presence of glucose, the Tt-7 transconjugants consumed glucose, leading to a better growth yield.

• Applied Biological Chemistry
• /
• v.41 no.5
• /
• pp.355-362
• /
• 1998

The minor form of phosphofructokinase (EC 2.7.1.11; PFK), which was suggested to be of plastid origin from the host fraction of chickpea nodules, was isolated as a small protein with apparent molecular mass near 220 kDa and purified to a high degree. SDS-PAGE and western blot indicated that the enzyme was made up of a homotetrameric structure (55 kDa). The enzyme had sharp pH profiles with maximal activities at pH 8 and displayed Michaelis-Menten kinetics with respect to Fru-6-P and nucleoside triphosphate substrate at the pH optimum (pH 8) and at pH 7. MgATP was the most effective phosphoryl donor. Phosphoenolpyruvate was a potent inhibitor of minor PFK activity, and the enzyme was also strongly inhibited by 3-phosphoglycerate, 2-phosphoglycerate, and to a lesser extent, PPi. Minor PFK was weakly activated by KCl, NaCl and Pi, and was inhibitory at high concentration of KCl and Pi.

• The Korean Journal of Pharmacology
• /
• v.28 no.1
• /
• pp.41-48
• /
• 1992

A concentration of 0.01 mM bupivacaine was necessary to maintain approximately 50% depression of contractility of rat atria suspended in a modified Krebs-Ringer bicarbonate glucose medium, pH 7.4 at $30^{\circ}C$. Sodium pyruvate, sodium acetate, and fructose partially restored the contractility of the bupivacaine-depressed atria. However, 20 mM glucose had no effect on the bupivacaine-depressed atria, although this concentration of glucose markedly increased the contractility of normal atria not to be exposed to bupivacaine. Contractility of normal atria was not significantly influenced by sodium pyruvate, sodium acetate, and fructose. The results suggested that at least part of the negative inotropic action of bupivacaine is the result of inhibition of glucose uptake or utilization in the glycolytic pathway, and further pinpoint the blockade as an early step in the glycolytic sequence prior to the phosphofructokinase step.

• Microbiology and Biotechnology Letters
• /
• v.24 no.3
• /
• pp.268-273
• /
• 1996

An extremely cadmium tolerant yeast, Hansenula anomala B-7 used to determine the modification of the intracellular enzyme activities by cadmium ion. The activities of alcohol dehydrogenase, phosphofructokinase, and cytidine deaminase were decreased up to 90%, 40%, and 86% compa- red with the control by 1 mM cadmium nitrate respectively, but the activities of malate dehydrogenase, 6- phosphogluconate dehydrogenase, cytochrome c oxidase, and alkaline phosphatase were increased up to 440%, 136%, 260% and 155% compared with the control by 1 mM cadmium nitrate respectively. These results show that the activities of the enzymes participating in Embden-Mayerhof pathway (e.g. anaerobic metabolism) were reduced by cadmium, but those involved in hexose monophosphate pathway and tricarboxylic acid cycle (e.g. aerobic metabolism) were stimulated in contrast. It has been suggested that the diminished activity of cytidine deaminase in pyrimidine nucleotide dissimilation occured due to the inhibited nucleotide dissimilation by cadmium ion; the enhanced activity of cytochrome c oxidase was specifically required in order to oxidize a raised amount of NADH and NADPH due to the increased aerobic metabolism.

• Journal of Ginseng Research
• /
• v.17 no.1
• /
• pp.13-21
• /
• 1993

We attempted in this study to understand the hypoglycemic action of the fat soluble fraction of red ginseng roots in streptozotocin injected diabetic rats, through its actions on several enzymes relating to carbohydrate metabolism of the 1eve1 to compare with those of ginsenosides in streptozotocin injected diabetic rats. It was realized that the increased level of glucose, ketone bodies, lactate, nonesterified fatty acids and triacylglycerol in blood was significantly decreased and the decreased liver glycogen content of streptozotocin injected rats were appreciably moderated by intraperitoneal injection of the fat soluble fraction of red ginseng roots as shown in the saponin injected diabetic rats. The deceased activities of liver enzymes relating to carbohydrate metabolism such as phosphofructokinase, glucokinase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and acetyl CoA carboxylase of streptozotocin induced diabetic rats were also sufficiently modified by the intraperitoneal injection of the above fat soluble fraction as shown in the ginsenoside injected streptozotocin induced rats.

• The Korean Journal of Pharmacology
• /
• v.31 no.1 s.57
• /
• pp.53-61
• /
• 1995

The experiments were performed to determine whether the cardiac depressant action of lidocaine is directly associated with the utilization of endogenous substrates in isolated rat atria, by using citrate and bicarbonate-free medium known as potent inhibitors of phosphofructokinases (PFK) enzyme step. Citrate and bicarbonate-free medium produced negative inotropic action of isolated rat atria incubated in normal Krebs-Ringer bicarbonate glucose medium. Pyruvate and acetate increased the force of contraction of atria depressed by citrate or bicarbonate-free medium, whereas fructose was without effect indicating the inhibitory effect of citrate and bicarbonate-free medium at some point in the glycolytic pathway such as the PFK step in atria. In the absence of exogenous substrate, citrate and bicarbonate-free medium produced a marked depression of the force of substrate-depleted atria indicating that utilization of endogenous substrate above the PFK step, probably cardiac glycogen, is also impaired by citrate or bicarbonate-free medium. Lidocaine produced further depression of the contractile force of atria depressed by citrate. These results argue strongly for an additional mechanism of cardiac depression caused by lidocaine involving the sites below the PFK.

• Journal of Life Science
• /
• v.29 no.11
• /
• pp.1179-1191
• /
• 2019

Cancer cells undergo the epithelial-mesenchymal transition (EMT) and show unique oncogenic metabolic phenotypes such as the glycolytic switch (Warburg effect) which are important for tumor development and progression. The EMT is a critical process for tumor invasion and metastasis. High-mobility group box 1 (HMGB1) is a chromatin-associated nuclear protein, but it acts as a damage-associated molecular pattern molecule when released from dying cells and immune cells. HMGB1 induces the EMT, as well as invasion and metastasis, thereby contributing to tumor progression. Here, we show that HMGB1 induced the EMT by activating Snail. In addition, the HMGB1/Snail cascade was found induce a glycolytic switch. HMGB1 also suppressed mitochondrial respiration and cytochrome c oxidase (COX) activity by a Snail-dependent reduction in the expression of the COX subunits COXVIIa and COXVIIc. HMGB1 also upregulated the expression of several key glycolytic enzymes, including hexokinase 2 (HK2), phosphofructokinase-2/fructose-2,6-bisphosphatase 2 (PFKFB2), and phosphoglycerate mutase 1 (PGAM1), in a Snail-dependent manner. However, HMGB1 was found to regulate some other glycolytic enzymes including lactate dehydrogenases A and B (LDHA and LDHB), glucose transporter 1 (GLUT1), and monocarboxylate transporters 1 and 4 (MCT1 and 4) in a Snail-independent manner. Transfection with short hairpin RNAs against HK2, PFKFB2, and PGAM1 prevented the HMGB1-induced EMT, indicating that glycolysis is associated with HMGB1-induced EMT. These findings demonstrate that HMGB1 signaling induces the EMT, glycolytic switch, and mitochondrial repression via Snail activation.

• The Korean Journal of Pharmacology
• /
• v.22 no.1 s.38
• /
• pp.51-59
• /
• 1986

The effect of metabolic substrate fructose on the force of contraction of isolated rat atria depressed with lidocaine was determined. Fructose produced dose-dependent increase in the force of contraction of isolated atria depressed by substrate-free Krebs-Ringer bicarbonate medium. The maximally effective concentration of fructose was 30 mM. The isolated atria, suspended in Krebs-Ringer bicarbonate glucose medium aerated with 95% $O_2-5%CO_2$at $30^{\circ}C$ and pH 7.4, were depressed 50% by approximately 2.34 mg/100 ml of lidocaine. Addition of 30 mM fructose to these depressed atria resulted in a marked increase in the contractile force similar to that with pyruvate and acetate. Fructose had no significant effect, however, on atria exposed to low-calcium medium. The results are consistent with a previous report suggesting blockade by lidocaine of the uptake or utilization of glucose in the glycolytic pathway, and further pinpoint the blockade as an early step in the glycolytic sequence prior to the phospho-fructokinase step.

• Journal of Chest Surgery
• /
• v.26 no.2
• /
• pp.75-79
• /
• 1993

Myocardial protection against ischemic and reperfusion injuries is still in troublesome eventhough couples of the way of myocardial protection have been applied since 1970's. One of the possibility in myocardial protection is adding Fructose-l,6-diphosphate(FDP) in cardioplegic solution. It is assumed that FDP can promote ATP production under anaerobic condition as well as inhibiting the supressing effect of lactate on phosphofructokinase. We compared the myocardial protecting effects of FDP in crystalloid cardioplegic solution (St. Thomas formula, 10$^{\circ}C$, pH = 7.4) and reperfusate using isolated rat hearts in modified Langendorf apparatus by the parameters of preischemic and post reperfusing heart rate, time to first beat, occurance of arrhythmia, time to stabilization, and the rate of left ventricular pressure developing. Group A (n = 10), containing no FDP in cardioplegic and reperfusing solutions was control. Group B (n = 5), containing FDP in cardioplegic solution, showed statistically significant superiority of postischemic left ventricular pressure development than the control group. Group C (n = 5), containing FDP in reperfusate, showed statistically significant myocardial depressing effect than the controls. Other parameters were unremarkable. The cause is uncertain, but it is assumed that the negative feedback inhibition of FDP in energy metabolism or unknown blocking effect of FDP on certain transmembrane ionic currents is present. In conclusion, 1) FDP in cardioplegic solution has beneficial effect on postischemic left ventricular preservation. 2) FDP is strong acid when is hydrolyzed, so precise acid titration is neccessary. 3) FDP in reperfusate has negative left ventricular preservation, otherwise the mechanism is still uncertain.