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Literature summary extracted from

  • Guerrero-Mendiola, C.; Garcia-Trejo, J.J.; Encalada, R.; Saavedra, E.; Ramirez-Silva, L.
    The contribution of two isozymes to the pyruvate kinase activity of Vibrio cholerae one K+-dependent constitutively active and another K+-independent with essential allosteric activation (2017), PLoS ONE, 12, e0178673 .
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.1.40 AMP slight activation Vibrio cholerae serotype O1
2.7.1.40 fructose 1,6-bisphosphate slight activation Vibrio cholerae serotype O1
2.7.1.40 fructose 1,6-bisphosphate slight activation, the isozyme with Glu117 is an active K+-dependent enzyme, at the same substrate concentration, its Vmax in the absence of fructose 1,6-bisphosphate is 80% of that with its effector. VcIPK is activated 31fold by fructose 1,6-bisphosphate Vibrio cholerae serotype O1
2.7.1.40 fructose 2,6-bisphosphate
-
 Vibrio cholerae serotype O1
2.7.1.40 fructose 6-phosphate slight activation Vibrio cholerae serotype O1
2.7.1.40 fructose 6-phosphate moderate activation Vibrio cholerae serotype O1
2.7.1.40 glucose 6-phosphate slight activation Vibrio cholerae serotype O1
2.7.1.40 glucose 6-phosphate VcIIPK is activated 159fold by glucose 6-phosphate Vibrio cholerae serotype O1
2.7.1.40 ribose 5-phosphate slight activation Vibrio cholerae serotype O1
2.7.1.40 ribose 5-phosphate Rib 5-P, VcIIPK is activated 200fold by ribose 5-phosphate. the pyruvate kinase with Lys117 is a K+-independent enzyme displaying an allosteric activation by ribose 5-phosphate. In the K+-independent enzyme, Mn2+ may mimic the allosteric effect of ribose 5-phosphate Vibrio cholerae serotype O1
2.7.1.40 ribose 5-phosphate the pyruvate kinase with Lys117 is a K+-independent enzyme displaying an allosteric activation by ribose 5-phosphate. In the K+-independent enzyme, Mn2+ may mimic the allosteric effect of ribose 5-phosphate Vibrio cholerae serotype O1

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.40 recombinant expression of His-tagged isozyme PKI from pMCSG7/VcIPK plasmid in Escherichia coli strain BL21-CodonPlus (DE3)-RIL Vibrio cholerae serotype O1
2.7.1.40 recombinant expression of His-tagged isozyme PKII from pMCSG7/VcIIPK plasmid in Escherichia coli strain BL21-CodonPlus (DE3)-RIL Vibrio cholerae serotype O1

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.40 ADP-Cr2+ a dead-end inhibitor; a dead-end inhibitor Vibrio cholerae serotype O1
2.7.1.40 oxalate a dead-end inhibitor; a dead-end inhibitor Vibrio cholerae serotype O1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.40 additional information
-
 additional information despite their different allosteric behavior, both isozymes display a rapid equilibrium random order kinetic mechanism. Kinetic analysis, detailed overview Vibrio cholerae serotype O1
2.7.1.40 0.06
-
 phosphoenolpyruvate pH 7.0, 25°C, recombinant non-His-tagged enzyme, in presence of fructose 1,6-bisphosphate Vibrio cholerae serotype O1
2.7.1.40 0.069
-
 phosphoenolpyruvate pH 7.0, 25°C, recombinant His-tagged enzyme, in presence of fructose 1,6-bisphosphate Vibrio cholerae serotype O1
2.7.1.40 0.26
-
 ADP pH 7.0, 25°C, recombinant His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 0.27
-
 ADP pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 0.55
-
 phosphoenolpyruvate pH 7.0, 25°C, recombinant His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 0.68
-
 phosphoenolpyruvate pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 1
-
 phosphoenolpyruvate pH 7.0, 25°C, recombinant His-tagged enzyme, in presence of glucose 6-phosphate Vibrio cholerae serotype O1

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.40 Cs+ activates, at saturating concentrations of PEP3-, ADP-Mg complex and free Mg2+, the Km for Cs+ is 13.5 mM Vibrio cholerae serotype O1
2.7.1.40 K+ K117 isozyme requires K+ Vibrio cholerae serotype O1
2.7.1.40 K+ K117 isozyme requires K+, at saturating concentrations of PEP3-, ADP-Mg complex and free Mg2+, the Km for K+ is 8.5 mM Vibrio cholerae serotype O1
2.7.1.40 Mg2+ required for activity Vibrio cholerae serotype O1
2.7.1.40 Mn2+ the K+-independent enzyme, Mn2+ may mimic the allosteric effect of ribose 5-phosphate Vibrio cholerae serotype O1
2.7.1.40 additional information the pyruvate kinase isozyme sequences with Glu117 have been found to be K+-dependent, whereas those with Lys117 are K+-independent Vibrio cholerae serotype O1
2.7.1.40 additional information the pyruvate kinase isozyme sequences with Glu117 have been found to be K+-dependent, whereas those with Lys117 are K+-independent. In comparison with other K+-dependent PKs, VcIPK exhibits higher kcat and 2-5fold lower Km for the monovalent cations. No activation by Na+ and Li+. VCIPK is a good example of type Ib activation by monovalent cations. In such a mechanism, M+ coordination is absolutely required for catalysis, where both kcat and kcat/Km increase hyperbolically with [K+] Vibrio cholerae serotype O1
2.7.1.40 NH4+ activates, at saturating concentrations of PEP3-, ADP-Mg complex and free Mg2+, the Km for NH4+ is 2.7 mM Vibrio cholerae serotype O1
2.7.1.40 Rb+ activates, at saturating concentrations of PEP3-, ADP-Mg complex and free Mg2+, the Km for Rb+ is 5.9 mM Vibrio cholerae serotype O1

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.40 200000
-
 recombinant His-tagged VcIIPK, native PAGE Vibrio cholerae serotype O1
2.7.1.40 200000
-
 recombinant His-tagged VcIPK, native PAGE Vibrio cholerae serotype O1

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.40 ADP + phosphoenolpyruvate Vibrio cholerae serotype O1
-
 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate Vibrio cholerae serotype O1 El Tor Inaba N16961
-
 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate Vibrio cholerae serotype O1 ATCC 39315
-
 ATP + pyruvate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.40 Vibrio cholerae serotype O1 Q9KQJ0
-
-
2.7.1.40 Vibrio cholerae serotype O1 Q9KUN0
-
-
2.7.1.40 Vibrio cholerae serotype O1 ATCC 39315 Q9KQJ0
-
-
2.7.1.40 Vibrio cholerae serotype O1 ATCC 39315 Q9KUN0
-
-
2.7.1.40 Vibrio cholerae serotype O1 El Tor Inaba N16961 Q9KQJ0
-
-
2.7.1.40 Vibrio cholerae serotype O1 El Tor Inaba N16961 Q9KUN0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.40 recombinant His-tagged isozyme PKI from Escherichia coli strain BL21-CodonPlus (DE3)-RIL by nickel affinity chromatography and ammonium sulfate fraction, the tag cannot be cleaved, because the cleavage site is not accessible to the TEV protease, to 90% purity Vibrio cholerae serotype O1
2.7.1.40 recombinant His-tagged isozyme PKII from Escherichia coli strain BL21-CodonPlus (DE3)-RIL by nickel affinity chromatography and ammonium sulfate fraction, the tag cannot be cleaved, because the cleavage site is not accessible to the TEV protease, to 88% purity Vibrio cholerae serotype O1

Source Tissue

EC Number Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.40 ADP + phosphoenolpyruvate
-
 Vibrio cholerae serotype O1 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate
-
 Vibrio cholerae serotype O1 El Tor Inaba N16961 ATP + pyruvate
-
 ?
2.7.1.40 ADP + phosphoenolpyruvate
-
 Vibrio cholerae serotype O1 ATCC 39315 ATP + pyruvate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.40 homotetramer 4 * 50000, recombinant His-tagged VcIIPK, SDS-PAGE, 4 * 54914, recombinant His-tagged VcIIPK, mass spectrometry, 4 * 52333, VcIIPK, sequence calculation Vibrio cholerae serotype O1
2.7.1.40 homotetramer 4 * 50000, recombinant His-tagged VcIPK, SDS-PAGE, 4 * 53138, recombinant His-tagged VcIPK, mass spectrometry, 4 * 50439, VcIPK, sequence calculation Vibrio cholerae serotype O1

Synonyms

EC Number Synonyms Comment Organism
2.7.1.40 K+-dependent PK
-
 Vibrio cholerae serotype O1
2.7.1.40 K+-independent PK
-
 Vibrio cholerae serotype O1
2.7.1.40 VcIIPK
-
 Vibrio cholerae serotype O1
2.7.1.40 VcIPK
-
 Vibrio cholerae serotype O1
2.7.1.40 VC_0485
-
 Vibrio cholerae serotype O1
2.7.1.40 VC_2008
-
 Vibrio cholerae serotype O1

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.40 25
-
 assay at Vibrio cholerae serotype O1

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.40 7
-
 assay at Vibrio cholerae serotype O1

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.7.1.40 0.0078
-
 oxalate pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 0.071
-
 oxalate pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 1.1
-
 ADP-Cr2+ pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1
2.7.1.40 1.3
-
 ADP-Cr2+ pH 7.0, 25°C, recombinant non-His-tagged enzyme Vibrio cholerae serotype O1

General Information

EC Number General Information Comment Organism
2.7.1.40 physiological function Vibrio cholerae has two isozymes that contribute to the pyruvate kinase activity: one K+-dependent constitutively active isozyme and another K+-independent isozyme with essential allosteric activation. The pyruvate kinase isozyme sequences with Glu117 have been found to be K+-dependent, whereas those with Lys117 are K+-independent Vibrio cholerae serotype O1