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P06999 (PFKB_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified July 9, 2014. Version 129. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (4) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Interactions·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
ATP-dependent 6-phosphofructokinase isozyme 2

Short name=ATP-PFK 2
Short name=Phosphofructokinase 2
EC=2.7.1.11
Alternative name(s):
6-phosphofructokinase isozyme II
Phosphohexokinase 2
Gene names
Name:pfkB
Ordered Locus Names:b1723, JW5280
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length309 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committing step of glycolysis. Ref.7

Catalytic activity

ATP + D-fructose 6-phosphate = ADP + D-fructose 1,6-bisphosphate. Ref.7 Ref.12

Cofactor

Magnesium. Ref.7

Enzyme regulation

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. Ref.7 Ref.12

Pathway

Carbohydrate degradation; glycolysis; D-glyceraldehyde 3-phosphate and glycerone phosphate from D-glucose: step 3/4.

Subunit structure

Homodimer.

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.

Sequence similarities

Belongs to the carbohydrate kinase PfkB family.

Biophysicochemical properties

Kinetic parameters:

KM=8 µM for ATP Ref.12

KM=6 µM for fructose 6-phosphate

Binary interactions

With

Entry

#Exp.

IntAct

Notes

itself2EBI-6966085,EBI-6966085

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 309309ATP-dependent 6-phosphofructokinase isozyme 2
PRO_0000080083

Regions

Nucleotide binding185 – 1873ATP
Nucleotide binding224 – 2296ATP
Region12 – 143Substrate binding
Region27 – 293Substrate binding
Region39 – 435Substrate binding
Region90 – 923Substrate binding
Region187 – 1893Allosteric inhibitor ATP binding

Sites

Active site2561 By similarity
Metal binding1901Magnesium; catalytic Probable
Metal binding2501Potassium; via carbonyl oxygen
Metal binding2521Potassium; via carbonyl oxygen
Metal binding2861Potassium; via carbonyl oxygen
Metal binding2891Potassium; via carbonyl oxygen
Metal binding2911Potassium; via carbonyl oxygen
Metal binding2931Potassium; via carbonyl oxygen
Binding site271Allosteric inhibitor ATP; shared with dimeric partner
Binding site271ATP; shared with dimeric partner
Binding site1391Substrate
Binding site2481ATP
Binding site2561Substrate
Binding site2801ATP; via carbonyl oxygen
Binding site2841ATP

Experimental info

Mutagenesis1901E → Q: Causes a 50-fold decrease in the kcat value and a 15-fold increment in the apparent KM for ATP. Ref.7
Sequence conflict26 – 3813GKLRC…VFEPG → ENCAVPHRCSNP in AAA24321. Ref.1
Sequence conflict26 – 3813GKLRC…VFEPG → ENCAVPHRCSNP in AAA24320. Ref.5
Sequence conflict155 – 17117AAQKQ…SSGEA → LRKNKGSAASSTVLGQG in AAA24321. Ref.1
Sequence conflict245 – 2462PV → AL in AAA24321. Ref.1
Sequence conflict257 – 2582SM → RL in AAA24321. Ref.1

Secondary structure

...................................................... 309
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P06999 [UniParc].

Last modified November 1, 1997. Version 2.
Checksum: A93BEBE0D5801309

FASTA30932,456
        10         20         30         40         50         60 
MVRIYTLTLA PSLDSATITP QIYPEGKLRC TAPVFEPGGG GINVARAIAH LGGSATAIFP 

        70         80         90        100        110        120 
AGGATGEHLV SLLADENVPV ATVEAKDWTR QNLHVHVEAS GEQYRFVMPG AALNEDEFRQ 

       130        140        150        160        170        180 
LEEQVLEIES GAILVISGSL PPGVKLEKLT QLISAAQKQG IRCIVDSSGE ALSAALAIGN 

       190        200        210        220        230        240 
IELVKPNQKE LSALVNRELT QPDDVRKAAQ EIVNSGKAKR VVVSLGPQGA LGVDSENCIQ 

       250        260        270        280        290        300 
VVPPPVKSQS TVGAGDSMVG AMTLKLAENA SLEEMVRFGV AAGSAATLNQ GTRLCSHDDT 


QKIYAYLSR 

« Hide

References

« Hide 'large scale' references
[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]"A 570-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 28.0-40.1 min region on the linkage map."
Aiba H., Baba T., Fujita K., Hayashi K., Inada T., Isono K., Itoh T., Kasai H., Kashimoto K., Kimura S., Kitakawa M., Kitagawa M., Makino K., Miki T., Mizobuchi K., Mori H., Mori T., Motomura K. expand/collapse author list , Nakade S., Nakamura Y., Nashimoto H., Nishio Y., Oshima T., Saito N., Sampei G., Seki Y., Sivasundaram S., Tagami H., Takeda J., Takemoto K., Takeuchi Y., Wada C., Yamamoto Y., Horiuchi T.
DNA Res. 3:363-377(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
[3]"The complete genome sequence of Escherichia coli K-12."
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.
Science 277:1453-1462(1997) [PubMed] [Europe PMC] [Abstract]
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.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
K02500 Genomic DNA. Translation: AAA24321.1.
U00096 Genomic DNA. Translation: AAC74793.1.
AP009048 Genomic DNA. Translation: BAA15500.2.
K00128 Genomic DNA. Translation: AAA24320.1.
PIRKIECFB. C64931.
RefSeqNP_416237.3. NC_000913.3.
YP_489984.1. NC_007779.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
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[»]
ProteinModelPortalP06999.
SMRP06999. Positions 1-309.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

DIPDIP-10465N.
MINTMINT-8178151.
STRING511145.b1723.

2D gel databases

SWISS-2DPAGEP06999.

Proteomic databases

PaxDbP06999.
PRIDEP06999.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC74793; AAC74793; b1723.
BAA15500; BAA15500; BAA15500.
GeneID12934082.
946230.
KEGGecj:Y75_p1698.
eco:b1723.
PATRIC32118753. VBIEscCol129921_1794.

Organism-specific databases

EchoBASEEB0694.
EcoGeneEG10700. pfkB.

Phylogenomic databases

eggNOGCOG1105.
HOGENOMHOG000265281.
KOK16370.
OMANVARTIH.
OrthoDBEOG61KBKF.
PhylomeDBP06999.

Enzyme and pathway databases

BioCycEcoCyc:6PFK-2-MONOMER.
ECOL316407:JW5280-MONOMER.
MetaCyc:6PFK-2-MONOMER.
SABIO-RKP06999.
UniPathwayUPA00109; UER00182.

Gene expression databases

GenevestigatorP06999.

Family and domain databases

Gene3D3.40.1190.20. 1 hit.
InterProIPR002173. Carboh/pur_kinase_PfkB_CS.
IPR011611. PfkB_dom.
IPR029056. Ribokinase-like.
IPR017583. Tagatose/fructose_Pkinase.
[Graphical view]
PfamPF00294. PfkB. 1 hit.
[Graphical view]
PIRSFPIRSF000535. 1PFK/6PFK/LacC. 1 hit.
SUPFAMSSF53613. SSF53613. 1 hit.
TIGRFAMsTIGR03168. 1-PFK. 1 hit.
PROSITEPS00583. PFKB_KINASES_1. 1 hit.
PS00584. PFKB_KINASES_2. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP06999.
PROP06999.

Entry information

Entry namePFKB_ECOLI
AccessionPrimary (citable) accession number: P06999
Secondary accession number(s): P78065, P78260
Entry history
Integrated into UniProtKB/Swiss-Prot: April 1, 1988
Last sequence update: November 1, 1997
Last modified: July 9, 2014
This is version 129 of the entry and version 2 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene