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Information on EC 2.7.1.146 - ADP-specific phosphofructokinase and Organism(s) Pyrococcus horikoshii and UniProt Accession O59355

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IUBMB Comments
ADP can be replaced by GDP, ATP and GTP, to a limited extent. Divalent cations are necessary for activity, with Mg2+ followed by Co2+ being the most effective.
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Pyrococcus horikoshii
UNIPROT: O59355
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The taxonomic range for the selected organisms is: Pyrococcus horikoshii
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
adp-dependent phosphofructokinase, tlpfk, tk0376, adp-dependent 6-phosphofructokinase, adp-dependent pfk, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ADP-dependent 6-phosphofructokinase
-
ADP-dependent phosphofructokinase
-
ADP dependent phosphofructokinase
-
-
-
-
ADP-6-phosphofructokinase
-
-
-
-
ADP-dependent 6-phosphofructokinase
-
-
ADP-dependent phosphofructokinase
-
-
-
-
ADP-Pfk
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
SYSTEMATIC NAME
IUBMB Comments
ADP:D-fructose-6-phosphate 1-phosphotransferase
ADP can be replaced by GDP, ATP and GTP, to a limited extent. Divalent cations are necessary for activity, with Mg2+ followed by Co2+ being the most effective.
CAS REGISTRY NUMBER
COMMENTARY hide
237739-62-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ADP + D-fructose 6-phosphate
AMP + D-fructose 1,6-bisphosphate
show the reaction diagram
-
-
-
?
D-fructose 6-phosphate + ADP
D-fructose 1,6-bisphosphate + AMP
show the reaction diagram
100% activity
-
-
?
D-fructose 6-phosphate + CDP
D-fructose 1,6-bisphosphate + CMP
show the reaction diagram
about 35% activity compared to ADP
-
-
?
D-fructose 6-phosphate + GDP
D-fructose 1,6-bisphosphate + GMP
show the reaction diagram
about 50% activity compared to ADP
-
-
?
D-fructose 6-phosphate + IDP
D-fructose 1,6-bisphosphate + IMP
show the reaction diagram
about 50% activity compared to ADP
-
-
?
D-fructose 6-phosphate + UDP
D-fructose 1,6-bisphosphate + UMP
show the reaction diagram
about 105% activity compared to ADP
-
-
?
ADP + D-fructose 6-phosphate
AMP + D-fructose 1,6-bisphosphate
show the reaction diagram
additional information
?
-
the PFK from Pyrococcus horikoshii (PhPFK) has no activity with glucose
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-
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ADP + D-fructose 6-phosphate
AMP + D-fructose 1,6-bisphosphate
show the reaction diagram
-
-
-
?
ADP + D-fructose 6-phosphate
AMP + D-fructose 1,6-bisphosphate
show the reaction diagram
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ni2+
highest activity in the presence of 2.5 mM NiSO4
CaCl2
2 mM ADP and 2 mM D-fructose 6-phosphate, CaCl2 show the lowest activity with 75% of the activity measured in the presence of MgCl2, activity does not change with the cation concentration in the range of 2.5 to 7 mM
MgSO4
2 mM ADP and 2 mM D-fructose 6-phosphate, magnesium is tested with 2 counter ions to discard any effect of the anion
MnCl2
2 mM ADP and 2 mM D-fructose 6-phosphate, increase in enzyme activity with the increase in MnCl2 concentration from 2.5 to 7 mM is observed
NiSO4
2 mM ADP and 2 mM D-fructose 6-phosphate, highest PhPFK activity is obtained with NiSO4 (97 units/mg), decrease in enzyme activity with the increase in MnCl2 concentration from 2.5 to 7 mM is observed
ZnCl2
2 mM ADP and 2 mM D-fructose 6-phosphate, no significant activity
additional information
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Ca2+
about 50% activity at 2.5 mM
Mg2+
about 60% activity at 2.5 mM
Mn2+
about 70% activity at 2.5 mM
NaCl
moderate inhibition by 300 mM NaCl or higher concentration
additional information
enzyme PhPFK is not significantly inhibited by D-glucose
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00747 - 0.473
ADP
0.00395 - 65
D-fructose 6-phosphate
0.00747 - 0.473
ADP
0.00395 - 22.6
D-fructose 6-phosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0048 - 151
D-fructose 6-phosphate
52 - 98
ADP
0.0048 - 151
D-fructose 6-phosphate
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.345 - 3830
ADP
0.0876 - 2990
D-fructose 6-phosphate
4400 - 8400
ADP
additional information
ADP
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
metabolism
Embden-Meyerhof glycolytic pathway, early step
additional information
molecular modeling, docking with D-glucose and D-fructose 6-phosphate, and molecular dynamics
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, the apo-enzyme is crystallized using 22% PEG 4000, 0.1 M Tris-HCl, pH 8.5, and 0.2 M LiSO4, at 21°C. Crystals of the PFK complex with AMP are obtained by crystallization of the PFK D17A protein in the presence of 20% PEG 3350, 0.2 M lithium citrate, 10 mM D-fructose 6-phosphate, and 5 mM ADP
through hanging drop vapor diffusion at 21°C by mixing protein solution with solution consisting of 22% PEG 4000, 0.1 M Tris-HCl, pH 8.5, and 0.2 M Li2SO4, crystals of the ADP-dependent 6-phosphofructokinase complex with AMP are obtained by crystallization of the mutant protein D17A protein in the presence of 20% PEG 3350, 0.2 M lithium citrate, 10 mM fructose 6-phosphate, and 5 mM ADPcrystal structure of ADP-dependent 6-phosphofructokinase in both apo- and AMP-bound forms determined to 2.0 A and 1.9 A resolution
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A71E
the mutant shows slightly decreased kcat compared to the wild type enzyme
D17A
the mutant shows severely decreased kcat compared to the wild type enzyme
D433A
the mutant shows no acitivity
K158A
the mutant shows slightly decreased kcat compared to the wild type enzyme
N15A
the mutant shows increased kcat compared to the wild type enzyme
N160A
the mutant shows strongly increased kcat compared to the wild type enzyme
Q224A
the mutant shows increased kcat compared to the wild type enzyme
Q97A
the mutant shows decreased kcat compared to the wild type enzyme
R185A
the mutant shows no acitivity
R191A
the mutant shows decreased kcat compared to the wild type enzyme
R191E
the mutant shows almost wild type enzyme activity
S183A
the mutant shows strongly increased kcat compared to the wild type enzyme
S183E
the mutant displays significantly less activity when the phosphoryl donor ADP is replaced with IDP, UDP, GDP, or CDP, compared with the wild type
S189A
the mutant shows decreased kcat compared to the wild type enzyme
S263A
the mutant shows increased kcat compared to the wild type enzyme
A71E
production by site-directed mutagenesis
D17A
production by site-directed mutagenesis
D433A
production by site-directed mutagenesis, loss of PFK activity
K158A
production by site-directed mutagenesis
N15A
production by site-directed mutagenesis
N160A
production by site-directed mutagenesis
Q224A
production by site-directed mutagenesis
Q97A
production by site-directed mutagenesis
R185A
production by site-directed mutagenesis
R191A
production by site-directed mutagenesis
R191E
production by site-directed mutagenesis
S183E
production by site-directed mutagenesis, the S183E mutant displays significantly less activity when the phosphoryl donor is replaced with IDP, UDP, GDP, or CDP compared with the wild type
S189A
production by site-directed mutagenesis
S263A
production by site-directed mutagenesis
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-NTA column chromatography
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, dialysis, and anion exchange chromatography
by using nickel-nitrilotriacetic acid affinity chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) and DL41(DE3) cells
gene pfkC, phylogenetic analysis, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
phylogenetic analysis
recombinant wild type and mutant native and selenomethionine-labeled proteins are expressed in BL21(DE3) and DL41(DE3) Escherichia coli cells
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Currie, M.A.; Merino, F.; Skarina, T.; Wong, A.H.; Singer, A.; Brown, G.; Savchenko, A.; Caniuguir, A.; Guixe, V.; Yakunin, A.F.; Jia, Z.
ADP-dependent 6-phosphofructokinase from Pyrococcus horikoshii OT3: structure determination and biochemical characterization of PH1645
J. Biol. Chem.
284
22664-22671
2009
Pyrococcus horikoshii, Pyrococcus horikoshii (O59355), Pyrococcus horikoshii OT-3 (O59355)
Manually annotated by BRENDA team
Merino, F.; Rivas-Pardo, J.A.; Caniuguir, A.; Garcia, I.; Guixe, V.
Catalytic and regulatory roles of divalent metal cations on the phosphoryl-transfer mechanism of ADP-dependent sugar kinases from hyperthermophilic archaea
Biochimie
94
516-524
2012
Pyrococcus horikoshii
Manually annotated by BRENDA team
Gonzalez-Ordenes, F.; Cea, P.A.; Fuentes-Ugarte, N.; Munoz, S.M.; Zamora, R.A.; Leonardo, D.; Garratt, R.C.; Castro-Fernandez, V.; Guixe, V.
ADP-dependent kinases from the archaeal order Methanosarcinales adapt to salt by a non-canonical evolutionarily conserved strategy
Front. Microbiol.
9
1305
2018
Methanohalobium evestigatum (D7E8P3), Methanohalobium evestigatum ATCC BAA-1072 (D7E8P3), Methanohalobium evestigatum DSM 3721 (D7E8P3), Methanohalobium evestigatum NBRC 107634 (D7E8P3), Methanohalobium evestigatum OCM 161 (D7E8P3), Methanohalobium evestigatum Z-7303 (D7E8P3), Methanosarcina mazei (Q8PZL9), Methanosarcina mazei ATCC BAA-159 (Q8PZL9), Methanosarcina mazei DSM 3647 (Q8PZL9), Methanosarcina mazei Goe1 (Q8PZL9), Methanosarcina mazei JCM 11833 (Q8PZL9), Methanosarcina mazei OCM 88 (Q8PZL9), Pyrococcus horikoshii (O59355), Pyrococcus horikoshii ATCC 700860 (O59355), uncultured bacterium
Manually annotated by BRENDA team
Castro-Fernandez, V.; Herrera-Morande, A.; Zamora, R.; Merino, F.; Gonzalez-Ordenes, F.; Padilla-Salinas, F.; Pereira, H.M.; Brandao-Neto, J.; Garratt, R.C.; Guixe, V.
Reconstructed ancestral enzymes reveal that negative selection drove the evolution of substrate specificity in ADP-dependent kinases
J. Biol. Chem.
292
15598-15610
2017
Methanocaldococcus jannaschii (Q58999), Pyrococcus horikoshii (O59355), Pyrococcus horikoshii ATCC 700860 (O59355), uncultured bacterium
Manually annotated by BRENDA team