Genomes and Genes
Gene Symbol: pfkB
Description: 6-phosphofructokinase II
Alias: ECK1721, JW5280
Species: Escherichia coli str. K-12 substr. MG1655
- Kotlarz D, Garreau H, Buc H. Regulation of the amount and of the activity of phosphofructokinases and pyruvate kinases in Escherichia coli. Biochim Biophys Acta. 1975;381:257-68 pubmed..Kinetics of depression after a shift to anaerobiosis have been followed and found to be of the order of the doubling time. ..
- Comer M. Gene organization around the phenylalanyl-transfer ribonucleic acid synthetase locus in Escherichia coli. J Bacteriol. 1981;146:269-74 pubmed..These deletions unambiguously defined the gene order as aroD pps himA pheT pheS thrS pfkB. Mutants with deletions covering either pheS or pheT, but not both, were analyzed further by assay of phenylalanyl-..
- Daldal F, Fraenkel D. Tn10 insertions in the pfkB region of Escherichia coli. J Bacteriol. 1981;147:935-43 pubmedThe locus pfkB is known to determine expression of a minor phosphofructokinase (Pfk-2). Pfk-2 and pfkB seem to be dispensable, since Tn10 insertions in pfkB, as well as deletions from Tn10 nearby, are obtainable...
- Kotlarz D, Buc H. Regulatory properties of phosphofructokinase 2 from Escherichia coli. Eur J Biochem. 1981;117:569-74 pubmed..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. ..
- Guixe V. Chemical modification of SH groups of E. coli phosphofructokinase-2 induces subunit dissociation: monomers are inactive but preserve ligand binding properties. Arch Biochem Biophys. 2000;376:313-9 pubmed..These results show the presence of SH groups in the interface of Pfk-2 subunits, critical for subunit interactions, and that conformational changes occurring through the dimers are essential for catalytic activity. ..
- Baez M, Cabrera R, Guixé V, Babul J. Unfolding pathway of the dimeric and tetrameric forms of phosphofructokinase-2 from Escherichia coli. Biochemistry. 2007;46:6141-8 pubmed
- Kirkpatrick C, Maurer L, Oyelakin N, Yoncheva Y, Maurer R, Slonczewski J. Acetate and formate stress: opposite responses in the proteome of Escherichia coli. J Bacteriol. 2001;183:6466-77 pubmed
- Vinopal R, Fraenkel D. PfkB and pfkC loci of Escherichia coli. J Bacteriol. 1975;122:1153-61 pubmed..We here describe a secondary mutation at pfkB, "PFKB-," which abolishes the suppression as well as the low residual activity of unsuppressed pfkA ..
- Guixe V, Babul J. Effect of ATP on phosphofructokinase-2 from Escherichia coli. A mutant enzyme altered in the allosteric site for MgATP. J Biol Chem. 1985;260:11001-5 pubmed..These results suggest the presence of an allosteric site for MgATP2- in Pfk-2 responsible for the MgATP2- inhibition, which is altered in Pfk-2* as a consequence of the structural mutation. ..
- Guixe V, Babul J. Influence of ligands on the aggregation of the normal and mutant forms of phosphofructokinase 2 of Escherichia coli. Arch Biochem Biophys. 1988;264:519-24 pubmed..Possibly Pfk-2* is not able to form a tetramer because of an alteration in the regulatory site for the Mg-nucleotide complex. ..
- Lengeler J. Analysis of mutations affecting the dissmilation of galactitol (dulcitol) in Escherichia coli K 12. Mol Gen Genet. 1977;152:83-91 pubmed
- Daldal F. Molecular cloning of the gene for phosphofructokinase-2 of Escherichia coli and the nature of a mutation, pfkB1, causing a high level of the enzyme. J Mol Biol. 1983;168:285-305 pubmedThe pfkB gene of Escherichia coli is known to specify a minor phosphofructokinase, Pfk-2, in the wild-type strain; the pfkB1 mutation causes a 25-fold increase in the amount of Pfk-2 so that it adequately substitutes for mutational loss ..
- Kim Y, Park J, Cho J, Cho K, Park Y, Lee J. Proteomic response analysis of a threonine-overproducing mutant of Escherichia coli. Biochem J. 2004;381:823-9 pubmed..coli mutant. ..
- Baez M, Babul J. Reversible unfolding of dimeric phosphofructokinase-2 from Escherichia coli reveals a dominant role of inter-subunit contacts for stability. FEBS Lett. 2009;583:2054-60 pubmed publisher..Pfk-2 undergoes a cooperative unfolding/dissociation process with the accumulation of an expanded and unstructured monomeric intermediate with a marginal stability and a large solvent accessibility with respect to the native dimer. ..
- Cabrera R, Ambrosio A, Garratt R, Guixé V, Babul J. Crystallographic structure of phosphofructokinase-2 from Escherichia coli in complex with two ATP molecules. Implications for substrate inhibition. J Mol Biol. 2008;383:588-602 pubmed publisher..This structure also provides the grounds to compare analogous features of the nonhomologous phosphofructokinases from E. coli. ..
- Guixe V, Rodriguez P, Babul J. Ligand-induced conformational transitions in Escherichia coli phosphofructokinase 2: evidence for an allosteric site for MgATP2-. Biochemistry. 1998;37:13269-75 pubmed..These results demonstrate the presence of an allosteric site for MgATP2- in Pfk-2 from E. coli, responsible for the inhibition of the enzyme activity by this ligand. ..
- Parducci R, Cabrera R, Baez M, Guixé V. Evidence for a catalytic Mg2+ ion and effect of phosphate on the activity of Escherichia coli phosphofructokinase-2: regulatory properties of a ribokinase family member. Biochemistry. 2006;45:9291-9 pubmed..Given that the E190Q mutant presents alterations in the inhibition by MgATP(2-) and phosphate, we conclude that the E190 residue has a role not only in catalysis but also in allosteric regulation. ..
- Baez M, Merino F, Astorga G, Babul J. Uncoupling the MgATP-induced inhibition and aggregation of Escherichia coli phosphofructokinase-2 by C-terminal mutations. FEBS Lett. 2008;582:1907-12 pubmed publisher..These results show that the D-T conversion could be uncoupled from the conformational changes that lead to the MgATP-induced allosteric inhibition. ..
- Babul J. Phosphofructokinases from Escherichia coli. Purification and characterization of the nonallosteric isozyme. J Biol Chem. 1978;253:4350-5 pubmed..e. pfkB+)...
- Cabrera R, Guixé V, Alfaro J, Rodriguez P, Babul J. Ligand-dependent structural changes and limited proteolysis of Escherichia coli phosphofructokinase-2. Arch Biochem Biophys. 2002;406:289-95 pubmed
- Thompson S, Cass K, Stellwagen E. Blue dextran-sepharose: an affinity column for the dinucleotide fold in proteins. Proc Natl Acad Sci U S A. 1975;72:669-72 pubmed..The procedure can be used to identify proteins, either purified or in crude cellular extracts, that possess the dinucleotide fold and to significantly improve the purification procedures for those proteins that possess the fold. ..
- Cabrera R, Babul J, Guixé V. Ribokinase family evolution and the role of conserved residues at the active site of the PfkB subfamily representative, Pfk-2 from Escherichia coli. Arch Biochem Biophys. 2010;502:23-30 pubmed publisher..Site-directed mutagenesis of R90 and D256 present in these motifs, indicate that R90 participates in the binding of the phosphorylated substrate and that D256 is involved in the phosphoryl transfer mechanism. ..
- Cabrera R, Caniuguir A, Ambrosio A, Guixé V, Garratt R, Babul J. Crystallization and preliminary crystallographic analysis of the tetrameric form of phosphofructokinase-2 from Escherichia coli, a member of the ribokinase family. Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006;62:935-7 pubmed..8, b = 86.8, c = 171.3 A. The calculated Matthews coefficient of 2.45 A(3) Da(-1) indicates the presence of two monomers in the asymmetric unit, corresponding to a solvent content of 49%. Structure determination is ongoing. ..
- Campos G, Guixe V, Babul J. Kinetic mechanism of phosphofructokinase-2 from Escherichia coli. A mutant enzyme with a different mechanism. J Biol Chem. 1984;259:6147-52 pubmed..For Pfk-2, ATP would be the first substrate to bind to the enzyme, and APD the last product to be released. ..
- Fenton A, Paricharttanakul N, Reinhart G. Disentangling the web of allosteric communication in a homotetramer: heterotropic activation in phosphofructokinase from Escherichia coli. Biochemistry. 2004;43:14104-10 pubmed..The free energies of the isolated interactions sum to 100% +/- 2% of the total. Therefore, the four unique interactions are all contributors to activation, are nonequivalent, and are additive. ..
- Peng L, Arauzo Bravo M, Shimizu K. Metabolic flux analysis for a ppc mutant Escherichia coli based on 13C-labelling experiments together with enzyme activity assays and intracellular metabolite measurements. FEMS Microbiol Lett. 2004;235:17-23 pubmed..9% of the carbon flux being channeled via the glyoxylate shunt. However, the flux of the pentose phosphate pathway significantly decreased in the ppc mutant. ..
- Fraenkel D, Kotlarz D, Buc H. Two fructose 6-phosphate kinase activities in Escherichia coli. J Biol Chem. 1973;248:4865-6 pubmed
- Baez M, Wilson C, Babul J. Folding kinetic pathway of phosphofructokinase-2 from Escherichia coli: a homodimeric enzyme with a complex domain organization. FEBS Lett. 2011;585:2158-64 pubmed publisher..We propose that these characteristics arise by a mutual constrain between the large domain and the ?-clasp domain imposed by their interrupted chain connectivity. ..