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American Journal of Hematology May 1982A case of M-type PFK deficiency due to the synthesis of a structurally abnormal and catalytically inactive M-subunit was reported. PFK activity was reduced (39% of...
A case of M-type PFK deficiency due to the synthesis of a structurally abnormal and catalytically inactive M-subunit was reported. PFK activity was reduced (39% of normal) in red cells, normal in leukocytes and platelets, and absent in muscle. The red cell enzyme was not inhibited by antiserum to human muscle PFK and displayed normal biochemical properties (Km for ATP and fructose-6-phosphate, storage stability at +4 degrees C and -80 degrees C, optimum pH, electrophoretic pattern and molecular weight). The complete lack of PFK activity in muscle was confirmed on both histological preparations and muscle extracts. Double immunodiffusion analysis using an antinormal M-PFK serum revealed that the enzyme molecule was present and immunologically identical with normal, although it was catalytically inactive. The muscle abnormality was also confirmed by electromyography, ischemic exercise testing, histochemistry and electron microscopy. Moreover, PFK activity was investigated in myoblast cultures maintained up to 25 days, and it was found to be absent.
Topics: Erythrocytes; Humans; Male; Middle Aged; Muscles; Phosphofructokinase-1
PubMed: 6211089
DOI: 10.1002/ajh.2830120303 -
Biochimie Jan 1984In this paper, we describe an efficient procedure for the purification of yeast phosphofructokinase. This procedure eliminates any time delay and enables to obtain an...
In this paper, we describe an efficient procedure for the purification of yeast phosphofructokinase. This procedure eliminates any time delay and enables to obtain an enzyme with minimum proteolytic alterations. The molecular weights of the oligomeric enzyme and of its constitutive subunits were both evaluated by means of several independent methods. However, the accuracy of each measurement was not sufficient to discriminate between an hexameric and an octameric structure of the enzyme oligomer. On the other hand, crosslinking experiments demonstrated the octameric structure of yeast phosphofructokinase. Obviously, some methods of molecular weight determination have led to erroneous results. In particular, our experiments show that the reliability of molecular weight determinations performed by gel filtration of native proteins must be considered with caution.
Topics: Cross-Linking Reagents; Macromolecular Substances; Molecular Weight; Phosphofructokinase-1; Protein Conformation; Saccharomyces cerevisiae; Scattering, Radiation; X-Rays
PubMed: 6231961
DOI: 10.1016/0300-9084(84)90191-3 -
Tanpakushitsu Kakusan Koso. Protein,... Apr 1968
Review
Topics: Adenosine Triphosphate; Amino Acids; Animals; Binding Sites; Catalysis; Cell Membrane Permeability; Citric Acid Cycle; Glycolysis; Hydrogen-Ion Concentration; Kinetics; Phosphofructokinase-1
PubMed: 4235600
DOI: No ID Found -
FEBS Letters Oct 1982
Topics: Glycolysis; Mutation; Phosphofructokinase-1; Saccharomyces cerevisiae
PubMed: 6217083
DOI: 10.1016/0014-5793(82)81053-3 -
Nature Communications Feb 2017Dynamic regulation of glucose flux between aerobic glycolysis and the pentose phosphate pathway (PPP) during epithelial-mesenchymal transition (EMT) is not...
Dynamic regulation of glucose flux between aerobic glycolysis and the pentose phosphate pathway (PPP) during epithelial-mesenchymal transition (EMT) is not well-understood. Here we show that Snail (SNAI1), a key transcriptional repressor of EMT, regulates glucose flux toward PPP, allowing cancer cell survival under metabolic stress. Mechanistically, Snail regulates glycolytic activity via repression of phosphofructokinase, platelet (PFKP), a major isoform of cancer-specific phosphofructokinase-1 (PFK-1), an enzyme involving the first rate-limiting step of glycolysis. The suppression of PFKP switches the glucose flux towards PPP, generating NADPH with increased metabolites of oxidative PPP. Functionally, dynamic regulation of PFKP significantly potentiates cancer cell survival under metabolic stress and increases metastatic capacities in vivo. Further, knockdown of PFKP rescues metabolic reprogramming and cell death induced by loss of Snail. Thus, the Snail-PFKP axis plays an important role in cancer cell survival via regulation of glucose flux between glycolysis and PPP.
Topics: Cell Survival; Epithelial-Mesenchymal Transition; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Glucose; Glycolysis; Humans; NADP; Neoplasms; Oxidative Stress; Pentose Phosphate Pathway; Phosphofructokinase-1; Phosphofructokinase-1, Type C; RNA, Small Interfering; Snail Family Transcription Factors
PubMed: 28176759
DOI: 10.1038/ncomms14374 -
The Biochemical Journal Apr 1990The control enzyme phosphofructokinase is of regulatory significance in the metabolism of glucose by the malarial parasite Plasmodium berghei. (1) The enzyme was...
The control enzyme phosphofructokinase is of regulatory significance in the metabolism of glucose by the malarial parasite Plasmodium berghei. (1) The enzyme was partially purified from erythrocytic stages of P. berghei by precipitation with poly(ethylene glycol) and chromatography on 2',5'-bisphosphoadenosine-Sepharose 4B. (2) Similarly to various other phosphofructokinases, the enzyme from P. berghei shows an allosteric behaviour. It is activated by fructose 6-phosphate and inhibited by ATP. (3) The effects of Mg2(+)-complexed ATP, free ATP and Mg2+ were studied by keeping constant the concentration of one of these and varying the concentrations of the other two. (4) The enzyme is shown to be allosterically inhibited by free ATP and by higher concentrations of Mg2+. Compared with phosphofructokinase of erythrocytes, inhibition by ATP is weaker by two orders of magnitude. Mg2(+)-complexed ATP has no effect on allosteric regulation. (5) The proposed kinetic model provides an adequate description of the data.
Topics: Adenosine Triphosphate; Animals; Kinetics; Magnesium; Mathematics; Models, Theoretical; Phosphofructokinase-1; Plasmodium berghei
PubMed: 2139776
DOI: 10.1042/bj2670353 -
Yeast (Chichester, England) Aug 2002Previously, studies on glucose-induced microautophagy in the methylotrophic yeast Pichia pastoris provided evidence that the glucose-induced selective...
Previously, studies on glucose-induced microautophagy in the methylotrophic yeast Pichia pastoris provided evidence that the glucose-induced selective autophagy-1-protein is the alpha-subunit of 6-phosphofructokinase (Pfk), a key enzyme in the glycolytic pathway. In our work, we could clearly demonstrate that two types of subunits of Pfk exist in P. pastoris. Investigating the yeast cell-free extract by Western blot analysis, two distinct signals of Pfk were obtained. In addition, we isolated a DNA sequence containing the complete ORF of PpPFK2 encoding the beta-subunit of Pfk from P. pastoris with a deduced molecular mass of 103.7 kDa. On the basis of these results, a hetero-oligomeric structure of Pfk in P. pastoris became obvious. Because the molecular and kinetic properties of a homo-oligomeric yeast Pfk appear to be more similar to those of mammalian Pfk, as described in the literature, our results are of interest for the growing number of studies on P. pastoris as a heterologous production system. Furthermore, the 3'- and 5'-non-coding regions of PpPFK2 were isolated and several putative binding sites for regulatory factors could be identified in the promoter region.
Topics: Amino Acid Sequence; Base Sequence; Blotting, Western; Cloning, Molecular; DNA, Fungal; Molecular Sequence Data; Open Reading Frames; Phosphofructokinase-1; Pichia; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid
PubMed: 12125051
DOI: 10.1002/yea.889 -
Methods in Enzymology 1975
Topics: 1-Propanol; Ammonium Sulfate; Chromatography, DEAE-Cellulose; Clostridium; Crystallization; Fractional Precipitation; Hot Temperature; Kinetics; Magnesium; Methods; Molecular Weight; Phosphofructokinase-1; Potassium; Spectrophotometry; Spectrum Analysis
PubMed: 124389
DOI: 10.1016/0076-6879(75)42098-5 -
Methods in Enzymology 1994
Comparative Study
Topics: Chromatography, Affinity; Chromatography, Gel; Coloring Agents; Dextrans; Indicators and Reagents; Kinetics; Osmolar Concentration; Phosphofructokinase-1; Polyethylene Glycols; Protein Binding; Saccharomyces cerevisiae; Salts; Solvents; Triazines
PubMed: 7519281
DOI: 10.1016/0076-6879(94)28015-0 -
Methods in Enzymology 1982
Comparative Study
Topics: Bacteria; Clostridium; Kinetics; Molecular Weight; NAD; Phosphofructokinase-1; Species Specificity; Spectrophotometry, Ultraviolet
PubMed: 6218378
DOI: 10.1016/s0076-6879(82)90110-0