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Protein

ATP-dependent 6-phosphofructokinase isozyme 2

Gene

pfkB

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committing step of glycolysis.1 Publication

Catalytic activityi

ATP + D-fructose 6-phosphate = ADP + D-fructose 1,6-bisphosphate.2 Publications

Cofactori

Mg2+1 Publication

Enzyme regulationi

Allosterically inhibited by ATP. Allosteric ATP-binding requires the presence of the substrate ATP. Inhibited by monovalent cations with ionic radii larger than Na+ (e.g. K+, Cs+). The monovalent cations increase the affinity of the allosteric site for ATP. PFK-2 is sensitive to inhibition by fructose 1,6-diphosphate.2 Publications

Kineticsi

  1. KM=8 µM for ATP1 Publication
  2. KM=6 µM for fructose 6-phosphate1 Publication

    Pathwayi

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei27 – 271Allosteric inhibitor ATP; shared with dimeric partner1 Publication
    Binding sitei27 – 271ATP; shared with dimeric partner3 Publications
    Binding sitei139 – 1391Substrate
    Metal bindingi190 – 1901Magnesium; catalyticCurated
    Binding sitei248 – 2481ATP3 Publications
    Metal bindingi250 – 2501Potassium; via carbonyl oxygen1 Publication
    Metal bindingi252 – 2521Potassium; via carbonyl oxygen1 Publication
    Active sitei256 – 2561By similarity
    Binding sitei256 – 2561Substrate
    Binding sitei280 – 2801ATP; via carbonyl oxygen3 Publications
    Binding sitei284 – 2841ATP3 Publications
    Metal bindingi286 – 2861Potassium; via carbonyl oxygen1 Publication
    Metal bindingi289 – 2891Potassium; via carbonyl oxygen1 Publication
    Metal bindingi291 – 2911Potassium; via carbonyl oxygen1 Publication
    Metal bindingi293 – 2931Potassium; via carbonyl oxygen1 Publication

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi185 – 1873ATP3 Publications
    Nucleotide bindingi224 – 2296ATP3 Publications

    GO - Molecular functioni

    • 6-phosphofructokinase activity Source: EcoCyc
    • ATP binding Source: UniProtKB-KW
    • identical protein binding Source: IntAct
    • magnesium ion binding Source: EcoCyc
    • tagatose-6-phosphate kinase activity Source: EcoCyc

    GO - Biological processi

    • carbohydrate phosphorylation Source: GOC
    • cellular response to DNA damage stimulus Source: EcoliWiki
    • glycolytic process Source: EcoCyc
    Complete GO annotation...

    Keywords - Molecular functioni

    Kinase, Transferase

    Keywords - Biological processi

    Glycolysis

    Keywords - Ligandi

    ATP-binding, Magnesium, Metal-binding, Nucleotide-binding

    Enzyme and pathway databases

    BioCyciEcoCyc:6PFK-2-MONOMER.
    ECOL316407:JW5280-MONOMER.
    MetaCyc:6PFK-2-MONOMER.
    BRENDAi2.7.1.11. 2026.
    SABIO-RKP06999.
    UniPathwayiUPA00109; UER00182.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    ATP-dependent 6-phosphofructokinase isozyme 2 (EC:2.7.1.11)
    Short name:
    ATP-PFK 2
    Short name:
    Phosphofructokinase 2
    Alternative name(s):
    6-phosphofructokinase isozyme II
    Phosphohexokinase 2
    Gene namesi
    Name:pfkB
    Ordered Locus Names:b1723, JW5280
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG10700. pfkB.

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi190 – 1901E → Q: Causes a 50-fold decrease in the kcat value and a 15-fold increment in the apparent KM for ATP. 1 Publication

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 309309ATP-dependent 6-phosphofructokinase isozyme 2PRO_0000080083Add
    BLAST

    Proteomic databases

    PaxDbiP06999.
    PRIDEiP06999.

    2D gel databases

    SWISS-2DPAGEP06999.

    Expressioni

    Gene expression databases

    GenevestigatoriP06999.

    Interactioni

    Subunit structurei

    Homodimer.5 Publications

    Binary interactionsi

    WithEntry#Exp.IntActNotes
    itself2EBI-6966085,EBI-6966085

    Protein-protein interaction databases

    DIPiDIP-10465N.
    MINTiMINT-8178151.
    STRINGi511145.b1723.

    Structurei

    Secondary structure

    1
    309
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Beta strandi4 – 74Combined sources
    Beta strandi12 – 209Combined sources
    Beta strandi24 – 296Combined sources
    Beta strandi34 – 396Combined sources
    Helixi40 – 5011Combined sources
    Beta strandi55 – 617Combined sources
    Helixi63 – 7513Combined sources
    Beta strandi80 – 845Combined sources
    Beta strandi93 – 975Combined sources
    Turni98 – 1003Combined sources
    Beta strandi103 – 1075Combined sources
    Helixi115 – 12511Combined sources
    Beta strandi133 – 1386Combined sources
    Helixi146 – 15813Combined sources
    Beta strandi162 – 1665Combined sources
    Helixi169 – 1757Combined sources
    Turni176 – 1783Combined sources
    Beta strandi181 – 1844Combined sources
    Helixi188 – 1958Combined sources
    Helixi204 – 21411Combined sources
    Beta strandi217 – 2193Combined sources
    Beta strandi221 – 2244Combined sources
    Helixi226 – 2283Combined sources
    Beta strandi230 – 2334Combined sources
    Beta strandi238 – 2414Combined sources
    Helixi254 – 26714Combined sources
    Helixi272 – 28615Combined sources
    Helixi297 – 30711Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3CQDX-ray1.98A/B1-309[»]
    3N1CX-ray2.00A/B/C/D1-309[»]
    3UMOX-ray1.70A/B1-309[»]
    3UMPX-ray1.85A/B1-309[»]
    3UQDX-ray2.14A/B/C/D1-309[»]
    3UQEX-ray2.20A/B1-309[»]
    ProteinModelPortaliP06999.
    SMRiP06999. Positions 1-309.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP06999.

    Family & Domainsi

    Region

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Regioni12 – 143Substrate binding
    Regioni27 – 293Substrate binding
    Regioni39 – 435Substrate binding
    Regioni90 – 923Substrate binding
    Regioni187 – 1893Allosteric inhibitor ATP binding

    Sequence similaritiesi

    Belongs to the carbohydrate kinase PfkB family.Curated

    Phylogenomic databases

    eggNOGiCOG1105.
    HOGENOMiHOG000265281.
    InParanoidiP06999.
    KOiK16370.
    OMAiDIAYQLD.
    OrthoDBiEOG61KBKF.
    PhylomeDBiP06999.

    Family and domain databases

    Gene3Di3.40.1190.20. 1 hit.
    InterProiIPR002173. Carboh/pur_kinase_PfkB_CS.
    IPR011611. PfkB_dom.
    IPR029056. Ribokinase-like.
    IPR017583. Tagatose/fructose_Pkinase.
    [Graphical view]
    PfamiPF00294. PfkB. 1 hit.
    [Graphical view]
    PIRSFiPIRSF000535. 1PFK/6PFK/LacC. 1 hit.
    SUPFAMiSSF53613. SSF53613. 1 hit.
    TIGRFAMsiTIGR03168. 1-PFK. 1 hit.
    PROSITEiPS00583. PFKB_KINASES_1. 1 hit.
    PS00584. PFKB_KINASES_2. 1 hit.
    [Graphical view]

    Sequencei

    Sequence statusi: Complete.

    P06999-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MVRIYTLTLA PSLDSATITP QIYPEGKLRC TAPVFEPGGG GINVARAIAH
    60 70 80 90 100
    LGGSATAIFP AGGATGEHLV SLLADENVPV ATVEAKDWTR QNLHVHVEAS
    110 120 130 140 150
    GEQYRFVMPG AALNEDEFRQ LEEQVLEIES GAILVISGSL PPGVKLEKLT
    160 170 180 190 200
    QLISAAQKQG IRCIVDSSGE ALSAALAIGN IELVKPNQKE LSALVNRELT
    210 220 230 240 250
    QPDDVRKAAQ EIVNSGKAKR VVVSLGPQGA LGVDSENCIQ VVPPPVKSQS
    260 270 280 290 300
    TVGAGDSMVG AMTLKLAENA SLEEMVRFGV AAGSAATLNQ GTRLCSHDDT

    QKIYAYLSR
    Length:309
    Mass (Da):32,456
    Last modified:November 1, 1997 - v2
    Checksum:iA93BEBE0D5801309
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti26 – 3813GKLRC…VFEPG → ENCAVPHRCSNP in AAA24321 (PubMed:6235149).CuratedAdd
    BLAST
    Sequence conflicti26 – 3813GKLRC…VFEPG → ENCAVPHRCSNP in AAA24320 (PubMed:6310120).CuratedAdd
    BLAST
    Sequence conflicti155 – 17117AAQKQ…SSGEA → LRKNKGSAASSTVLGQG in AAA24321 (PubMed:6235149).CuratedAdd
    BLAST
    Sequence conflicti245 – 2462PV → AL in AAA24321 (PubMed:6235149).Curated
    Sequence conflicti257 – 2582SM → RL in AAA24321 (PubMed:6235149).Curated

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    K02500 Genomic DNA. Translation: AAA24321.1.
    U00096 Genomic DNA. Translation: AAC74793.1.
    AP009048 Genomic DNA. Translation: BAA15500.2.
    K00128 Genomic DNA. Translation: AAA24320.1.
    PIRiC64931. KIECFB.
    RefSeqiNP_416237.3. NC_000913.3.

    Genome annotation databases

    EnsemblBacteriaiAAC74793; AAC74793; b1723.
    BAA15500; BAA15500; BAA15500.
    GeneIDi946230.
    KEGGiecj:Y75_p1698.
    eco:b1723.
    PATRICi32118753. VBIEscCol129921_1794.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    K02500 Genomic DNA. Translation: AAA24321.1.
    U00096 Genomic DNA. Translation: AAC74793.1.
    AP009048 Genomic DNA. Translation: BAA15500.2.
    K00128 Genomic DNA. Translation: AAA24320.1.
    PIRiC64931. KIECFB.
    RefSeqiNP_416237.3. NC_000913.3.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    3CQDX-ray1.98A/B1-309[»]
    3N1CX-ray2.00A/B/C/D1-309[»]
    3UMOX-ray1.70A/B1-309[»]
    3UMPX-ray1.85A/B1-309[»]
    3UQDX-ray2.14A/B/C/D1-309[»]
    3UQEX-ray2.20A/B1-309[»]
    ProteinModelPortaliP06999.
    SMRiP06999. Positions 1-309.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-10465N.
    MINTiMINT-8178151.
    STRINGi511145.b1723.

    2D gel databases

    SWISS-2DPAGEP06999.

    Proteomic databases

    PaxDbiP06999.
    PRIDEiP06999.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC74793; AAC74793; b1723.
    BAA15500; BAA15500; BAA15500.
    GeneIDi946230.
    KEGGiecj:Y75_p1698.
    eco:b1723.
    PATRICi32118753. VBIEscCol129921_1794.

    Organism-specific databases

    EchoBASEiEB0694.
    EcoGeneiEG10700. pfkB.

    Phylogenomic databases

    eggNOGiCOG1105.
    HOGENOMiHOG000265281.
    InParanoidiP06999.
    KOiK16370.
    OMAiDIAYQLD.
    OrthoDBiEOG61KBKF.
    PhylomeDBiP06999.

    Enzyme and pathway databases

    UniPathwayiUPA00109; UER00182.
    BioCyciEcoCyc:6PFK-2-MONOMER.
    ECOL316407:JW5280-MONOMER.
    MetaCyc:6PFK-2-MONOMER.
    BRENDAi2.7.1.11. 2026.
    SABIO-RKP06999.

    Miscellaneous databases

    EvolutionaryTraceiP06999.
    PROiP06999.

    Gene expression databases

    GenevestigatoriP06999.

    Family and domain databases

    Gene3Di3.40.1190.20. 1 hit.
    InterProiIPR002173. Carboh/pur_kinase_PfkB_CS.
    IPR011611. PfkB_dom.
    IPR029056. Ribokinase-like.
    IPR017583. Tagatose/fructose_Pkinase.
    [Graphical view]
    PfamiPF00294. PfkB. 1 hit.
    [Graphical view]
    PIRSFiPIRSF000535. 1PFK/6PFK/LacC. 1 hit.
    SUPFAMiSSF53613. SSF53613. 1 hit.
    TIGRFAMsiTIGR03168. 1-PFK. 1 hit.
    PROSITEiPS00583. PFKB_KINASES_1. 1 hit.
    PS00584. PFKB_KINASES_2. 1 hit.
    [Graphical view]
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Nucleotide sequence of gene pfkB encoding the minor phosphofructokinase of Escherichia coli K-12."
      Daldal F.
      Gene 28:337-342(1984) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: K12.
    2. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    3. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    4. "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
      Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
      Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    5. "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."
      Daldal F.
      J. Mol. Biol. 168:285-305(1983) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-38.
    6. "Escherichia coli proteome analysis using the gene-protein database."
      VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.
      Electrophoresis 18:1243-1251(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: IDENTIFICATION BY 2D-GEL.
    7. "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."
      Parducci R.E., Cabrera R., Baez M., Guixe V.
      Biochemistry 45:9291-9299(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: FUNCTION, CATALYTIC ACTIVITY, COFACTOR, MUTAGENESIS OF GLU-190, ENZYME REGULATION.
    8. "Crystallographic structure of phosphofructokinase-2 from Escherichia coli in complex with two ATP molecules. Implications for substrate inhibition."
      Cabrera R., Ambrosio A.L., Garratt R.C., Guixe V., Babul J.
      J. Mol. Biol. 383:588-602(2008) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.98 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ATP AND ALLOSTERIC INHIBITOR ATP.
    9. "The crystal complex of phosphofructokinase-2 of Escherichia coli with fructose-6-phosphate: kinetic and structural analysis of the allosteric ATP inhibition."
      Cabrera R., Baez M., Pereira H.M., Caniuguir A., Garratt R.C., Babul J.
      J. Biol. Chem. 286:5774-5783(2011) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEX WITH FRUCTOSE-6-PHOSPHATE.
    10. "Structure of E. coli PFK2 in complex with substrates and products."
      Pereira H.M., Caniuguir A., Baez M., Cabrera R., Garratt R.C., Babul J.
      Submitted (NOV-2011) to the PDB data bank
      Cited for: X-RAY CRYSTALLOGRAPHY (2.14 ANGSTROMS) IN COMPLEX WITH ADP; ATP; FRUCTOSE-1-6-DIPHOSPHATE AND FRUCTOSE-6-PHOSPHATE.
    11. "Structure of E. coli PFK2 mutant Y23D."
      Pereira H.M., Caniuguir A., Baez M., Cabrera R., Garratt R.C., Babul J.
      Submitted (NOV-2011) to the PDB data bank
      Cited for: X-RAY CRYSTALLOGRAPHY (2.20 ANGSTROMS) IN COMPLEX WITH ATP AND DIPHOSPHATE.
    12. "A ribokinase family conserved monovalent cation binding site enhances the MgATP-induced inhibition in E. coli phosphofructokinase-2."
      Baez M., Cabrera R., Pereira H.M., Blanco A., Villalobos P., Ramirez-Sarmiento C.A., Caniuguir A., Guixe V., Garratt R.C., Babul J.
      Biophys. J. 105:185-193(2013) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEX WITH ATP AND POTASSIUM, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION.

    Entry informationi

    Entry nameiPFKB_ECOLI
    AccessioniPrimary (citable) accession number: P06999
    Secondary accession number(s): P78065, P78260
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: April 1, 1988
    Last sequence update: November 1, 1997
    Last modified: May 27, 2015
    This is version 136 of the entry and version 2 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Miscellaneous

    Only 10% of the activity present in the wild-type strain is phosphofructokinase-2.
    This enzyme is not to be confused with 6-phosphofructo-2-kinase which is also called phosphofructokinase 2.
    E.coli has two 6-phosphofructokinases enzymes.

    Keywords - Technical termi

    3D-structure, Complete proteome, Reference proteome

    Documents

    1. Escherichia coli
      Escherichia coli (strain K12): entries and cross-references to EcoGene
    2. PATHWAY comments
      Index of metabolic and biosynthesis pathways
    3. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    4. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.