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Cell Reports Oct 2023Metastasis is the leading cause of high ovarian-cancer-related mortality worldwide. Three major processes constitute the whole metastatic cascade: invasion,...
Metastasis is the leading cause of high ovarian-cancer-related mortality worldwide. Three major processes constitute the whole metastatic cascade: invasion, intravasation, and extravasation. Tumor cells often reprogram their metabolism to gain advantages in proliferation and survival. However, whether and how those metabolic alterations contribute to the invasiveness of tumor cells has yet to be fully understood. Here we performed a genome-wide CRISPR-Cas9 screening to identify genes participating in tumor cell dissemination and revealed that PTGES3 acts as an invasion suppressor in ovarian cancer. Mechanistically, PTGES3 binds to phosphofructokinase, liver type (PFKL) and generates a local source of prostaglandin E2 (PGE2) to allosterically inhibit the enzymatic activity of PFKL. Repressed PFKL leads to downgraded glycolysis and the subsequent TCA cycle for glucose metabolism. However, ovarian cancer suppresses the expression of PTGES3 and disrupts the PTGES3-PGE2-PFKL inhibitory axis, leading to hyperactivation of glucose oxidation, eventually facilitating ovarian cancer cell motility and invasiveness.
Topics: Humans; Female; Dinoprostone; Phosphofructokinases; Phosphofructokinase-1; Liver; Glucose; Ovarian Neoplasms; Cell Proliferation; Cell Line, Tumor; Neoplasm Invasiveness
PubMed: 37831605
DOI: 10.1016/j.celrep.2023.113246 -
Investigative Ophthalmology & Visual... Nov 1984The activity of phosphofructokinase (PFK), a key regulatory enzyme of glycolysis, has been measured in the 27,000 X g supernatant of homogenates prepared from excised...
The activity of phosphofructokinase (PFK), a key regulatory enzyme of glycolysis, has been measured in the 27,000 X g supernatant of homogenates prepared from excised calf trabecular meshwork. The enzyme required NH4+, both at pH 8.5 and pH 7.2. This requirement was not relieved by K+ or AMP. At pH 7.2 and ATP levels of 0.1 to 2.5 mM, PFK was completely inactive in the absence of added AMP or NH4+. PFK was only weakly activated by 0.5 mM AMP or by 5 mM NH4+, but in the presence of both AMP and NH4+, PFK was highly active up to 1 mM ATP. At pH 8.5 and ATP levels of 0.1-12.5 mM, PFK was weakly active in the absence of added NH4+, with or without AMP. With the addition of 5 mM NH4+, PFK was highly active up to 2.5 mM ATP, while AMP was largely without effect. Concentrations of NH4+ as low as 0.03 mM stimulated PFK activity to 20% of maximal, yet the maximum was not reached until NH4+ levels were 10-30 mM. The activation of PFK by AMP and its inhibition by ATP is profoundly modified by pH. In contrast, the requirement for NH4+ is unaffected. This requirement suggests a regulatory role for ammonium ion in controlling the rate of glycolysis in trabecular meshwork. The concentration of ammonium in calf aqueous humor was found to be 0.18 mM, which is in the right range to have an effect.
Topics: Animals; Cattle; Enzyme Activation; Hydrogen-Ion Concentration; Phosphofructokinase-1; Quaternary Ammonium Compounds; Trabecular Meshwork
PubMed: 6238014
DOI: No ID Found -
Journal of Cellular Biochemistry May 2012Kinetic analysis of PFK-1 from rodent AS-30D, and human HeLa and MCF-7 carcinomas revealed sigmoidal [fructose 6-phosphate, Fru6P]-rate curves with different V(m) values...
Kinetic analysis of PFK-1 from rodent AS-30D, and human HeLa and MCF-7 carcinomas revealed sigmoidal [fructose 6-phosphate, Fru6P]-rate curves with different V(m) values when varying the allosteric activator fructose 2,6 bisphosphate (Fru2,6BP), AMP, Pi, NH(4)(+), or K(+). The rate equation that accurately predicted this behavior was the exclusive ligand binding concerted transition model together with non-essential hyperbolic activation. PFK-1 from rat liver and heart also exhibited the mixed cooperative-hyperbolic kinetic behavior regarding activators. Lowering pH induced decreased affinity for Fru6P, Fru2,6BP, citrate, and ATP (as inhibitor); as well as decreased V(m) and increased content of inactive (T) enzyme forms. High K(+) prompted increased (Fru6P) or decreased (activators) affinities; increased V(m); and increased content of active (R) enzyme forms. mRNA expression analysis and nucleotide sequencing showed that the three PFK-1 isoforms L, M, and C are transcribed in the three carcinomas. However, proteomic analysis indicated the predominant expression of L in liver, of M in heart and MCF-7 cells, of L>M in AS-30D cells, and of C in HeLa cells. PFK-1M showed the highest affinities for F6P and citrate and the lowest for ATP (substrate) and F2,6BP; PFK-1L showed the lowest affinity for F6P and the highest for F2,6BP; and PFK-1C exhibited the highest affinity for ATP (substrate) and the lowest for citrate. Thus, the present work documents the kinetic signature of each PFK-1 isoform, and facilitates the understanding of why this enzyme exerts significant or negligible glycolysis flux-control in normal or cancer cells, respectively, and how it regulates the onset of the Pasteur effect.
Topics: Animals; Base Sequence; Breast Neoplasms; Cell Line, Tumor; DNA, Complementary; Enzyme Activation; Female; HeLa Cells; Humans; Kinetics; Liver; Liver Neoplasms, Experimental; Myocardium; Neoplasms; Phosphofructokinase-1; Phosphofructokinase-1, Liver Type; Phosphofructokinase-1, Muscle Type; Phosphofructokinase-1, Type C; Polymorphism, Genetic; Rats; Rats, Wistar; Substrate Specificity; Uterine Cervical Neoplasms
PubMed: 22213537
DOI: 10.1002/jcb.24039 -
Biochemical and Biophysical Research... May 1965
Topics: Erythrocytes; Genetics, Medical; Glycogen Storage Disease; Glycogen Storage Disease Type VII; Glycolysis; Humans; Lactates; Muscle, Skeletal; Muscles; Phosphofructokinase-1; Phosphofructokinases; Physical Exertion
PubMed: 14339001
DOI: 10.1016/0006-291x(65)90156-7 -
Methods in Enzymology 1975
Topics: 1-Propanol; Ammonium Sulfate; Animals; Crystallization; Fractional Precipitation; Freezing; Hot Temperature; Methods; Muscles; Phosphofructokinase-1; Rabbits; Spectrophotometry
PubMed: 124387
DOI: 10.1016/0076-6879(75)42096-1 -
Methods in Enzymology 1982
Topics: Animals; Carcinoma, Ehrlich Tumor; Kinetics; Mice; Mice, Inbred Strains; NAD; Phosphofructokinase-1; Spectrophotometry, Ultraviolet
PubMed: 6218371
DOI: 10.1016/s0076-6879(82)90103-3 -
The Journal of Biological Chemistry Oct 1972
Topics: Adenosine Monophosphate; Adenosine Triphosphate; Allosteric Regulation; Animals; Caffeine; Charcoal; Cyclic AMP; Enzyme Activation; Epinephrine; Fructosephosphates; Glucosephosphates; Hydrogen-Ion Concentration; Male; Muscles; Myocardium; Phosphofructokinase-1; Protein Binding; Rabbits; Sheep
PubMed: 4346801
DOI: No ID Found -
Biochemical and Biophysical Research... Aug 1981
Topics: Adenosine Triphosphate; Animals; Citrates; Citric Acid; Fructosediphosphates; Fructosephosphates; Hexosediphosphates; Kinetics; Liver; Phosphates; Phosphofructokinase-1; Rats
PubMed: 6458291
DOI: 10.1016/0006-291x(81)91859-3 -
European Journal of Biochemistry Jul 1981Escherichia coli K12 contains two phosphofructokinases: phosphofructokinase 1, the most studied one, appears to behave as an allosteric enzyme, while phosphofructokinase...
Escherichia coli K12 contains two phosphofructokinases: phosphofructokinase 1, the most studied one, appears to behave as an allosteric enzyme, while phosphofructokinase 2 presents the features of a Michaelian enzyme. We show the present paper that, in fact, phosphofructokinase 2 also presents some regulatory properties in vitro: at high concentrations, ATP is an inhibitor of phosphofructokinase 2 and it provokes the tetramerization of the dimeric native enzyme. The binding of the two substrates to phosphofructokinase 2 is sequential and ordered as for phosphofructokinase 1, but in the former case fructose 6-phosphate is the first substrate to be bound and ADP the first product to be released. Each dimer of phosphofructokinase 2 binds two molecules of fructose 6-phosphate but only one molecule of the product fructose 1,6-phosphate. Although both phosphofructokinases of E. coli K12 present regulatory properties in vitro, the mechanism of regulation of the activity of the two enzymes is strikingly different. It can be asked whether or not these mechanisms operate in vivo.
Topics: Escherichia coli; Isoenzymes; Kinetics; Macromolecular Substances; Phosphofructokinase-1; Species Specificity
PubMed: 6456900
DOI: 10.1111/j.1432-1033.1981.tb06375.x -
Archives of Biochemistry and Biophysics Aug 1976
Topics: Adenine Nucleotides; Animals; Binding Sites; Binding, Competitive; Chromatography, Affinity; Inosine Nucleotides; Muscles; Phosphofructokinase-1; Protein Binding; Rabbits
PubMed: 134285
DOI: 10.1016/0003-9861(76)90536-1