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Literature summary extracted from

  • Zoraghi, R.; See, R.H.; Gong, H.; Lian, T.; Swayze, R.; Finlay, B.B.; Brunham, R.C.; McMaster, W.R.; Reiner, N.E.
    Functional analysis, overexpression, and kinetic characterization of pyruvate kinase from methicillin-resistant Staphylococcus aureus (2010), Biochemistry, 49, 7733-7747.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.7.1.40 AMP allosteric activator Staphylococcus aureus
2.7.1.40 ATP slight activation of the full-length enzyme, not the C-terminally truncated enzyme Staphylococcus aureus
2.7.1.40 D-ribose 5-phosphate allosteric activator Staphylococcus aureus
2.7.1.40 additional information no activation by fructose 1,6-bisphosphate Staphylococcus aureus

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.1.40 gene pyk, phylogenetic tree, expression of His-tagged full-length and C-terminally truncated enzymes in Escherichia coli strain BL-21(DE3) Staphylococcus aureus
2.7.1.40 gene pykA, phylogenetic tree, expression of His-tagged isozyme PK2 in Escherichia coli strain BL21(DE3) Escherichia coli
2.7.1.40 gene pykf, phylogenetic tree, expression of His-tagged isozyme PK1 in Escherichia coli strain BL21(DE3) Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.7.1.40 additional information construction of a C-terminally truncated mutant PKCT with a stop after residue 390. The catalytic activities of PKCT toward both phophoenolpyruvate and ADP are profoundly decreased compared to those of wild-type enzyme Staphylococcus aureus

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.7.1.40 ATP inhibition of full-length enzyme at concentration above 2.5 mM Staphylococcus aureus
2.7.1.40 D-fructose 1,6-bisphosphate inhibition of full-length enzyme at 10 mM Staphylococcus aureus
2.7.1.40 phosphate
-
 Staphylococcus aureus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.1.40 additional information
-
 additional information steady-state kinetics of recombinant wild-type and truncated PKin the absence or presence of 1 mM AMP as a function of phosphoenolpyruvate Staphylococcus aureus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.1.40 Mg2+ required Staphylococcus aureus
2.7.1.40 Mg2+ required Escherichia coli

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.1.40 53100
-
 4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation Staphylococcus aureus
2.7.1.40 65100
-
 4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation Staphylococcus aureus
2.7.1.40 215000
-
 His-tagged C-terminally truncated enzyme mutant, gel filtration Staphylococcus aureus
2.7.1.40 250000
-
 His-tagged full-length enzyme, gel filtration Staphylococcus aureus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.1.40 ATP + pyruvate Staphylococcus aureus
-
 ADP + phosphoenolpyruvate
-
 ?
2.7.1.40 ATP + pyruvate Escherichia coli
-
 ADP + phosphoenolpyruvate
-
 ?
2.7.1.40 ATP + pyruvate Staphylococcus aureus MRSA252
-
 ADP + phosphoenolpyruvate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.1.40 Escherichia coli A0A0H3JER5 isozyme PK2; gene pykA
-
2.7.1.40 Escherichia coli P0AD62 isozyme PK1; gene pykF
-
2.7.1.40 Staphylococcus aureus Q6GG09 a methicillin-resistant strain, gene pyk
-
2.7.1.40 Staphylococcus aureus MRSA252 Q6GG09 a methicillin-resistant strain, gene pyk
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.1.40 recombinant His-tagged full-length and C-terminally truncated enzymes from Escherichia coli strain BL-21(DE3) by nickel affiniy chromatography Staphylococcus aureus
2.7.1.40 recombinant His-tagged isozyme PK1 from Escherichia coli strain BL21(DE3) by nickel affiniy chromatography Escherichia coli
2.7.1.40 recombinant His-tagged isozyme PK2 from Escherichia coli strain BL21(DE3) by nickel affiniy chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.1.40 ATP + pyruvate
-
 Staphylococcus aureus ADP + phosphoenolpyruvate
-
 ?
2.7.1.40 ATP + pyruvate
-
 Escherichia coli ADP + phosphoenolpyruvate
-
 ?
2.7.1.40 ATP + pyruvate
-
 Staphylococcus aureus MRSA252 ADP + phosphoenolpyruvate
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.1.40 homotetramer
-
 Escherichia coli
2.7.1.40 homotetramer 4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation Staphylococcus aureus
2.7.1.40 More the C-terminal domain is not required for the tetramerization of the enzyme, homotetramerization also occurs in a truncated enzyme lacking the domain Staphylococcus aureus

Synonyms

EC Number Synonyms Comment Organism
2.7.1.40 MRSA PK
-
 Staphylococcus aureus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.1.40 37
-
 assay at Staphylococcus aureus
2.7.1.40 37
-
 assay at Escherichia coli

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.1.40 7.5
-
 assay at Staphylococcus aureus
2.7.1.40 7.5
-
 assay at Escherichia coli

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.1.40 ATP
-
 Staphylococcus aureus
2.7.1.40 ATP
-
 Escherichia coli

General Information

EC Number General Information Comment Organism
2.7.1.40 evolution significant evolutionary distance existing between the type I and type II isoenzymes in Gram-negative bacteria Staphylococcus aureus
2.7.1.40 malfunction the catalytic activities of the C-terminally truncated mutant toward both phophoenolpyruvate and ADP are profoundly decreased compared to those of wild-type enzyme Staphylococcus aureus
2.7.1.40 additional information the C-terminally truncated enzyme exhibits high affinity toward both phophoenolpyruvate and ADP and exhibits hyperbolic kinetics toward phophoenolpyruvate in the presence of activators AMP and ribose 5-phosphate consistent with kinetic properties of full-length enzyme Staphylococcus aureus
2.7.1.40 physiological function the C-terminal domain is not required for substrate binding or allosteric regulation observed in the holoenzyme, the kinetic efficiency of the truncated enzyme is decreased by 24 and 16fold, in ligand-free state, toward phophoenolpyruvate and ADP, respectively, but is restored by 3fold in AMP-bound state. The C-terminal domain (Gly473-Leu585) plays a substantial role in enzyme activity and comformational stability, and the C-terminal domain is involved in maintaining the specificity of allosteric regulation Staphylococcus aureus