Activating Compound | Comment | Organism | Structure |
---|---|---|---|
AMP | allosteric activator | Staphylococcus aureus | |
ATP | slight activation of the full-length enzyme, not the C-terminally truncated enzyme | Staphylococcus aureus | |
D-ribose 5-phosphate | allosteric activator | Staphylococcus aureus | |
additional information | no activation by fructose 1,6-bisphosphate | Staphylococcus aureus |
Cloned (Comment) | Organism |
---|---|
gene pyk, phylogenetic tree, expression of His-tagged full-length and C-terminally truncated enzymes in Escherichia coli strain BL-21(DE3) | Staphylococcus aureus |
gene pykA, phylogenetic tree, expression of His-tagged isozyme PK2 in Escherichia coli strain BL21(DE3) | Escherichia coli |
gene pykf, phylogenetic tree, expression of His-tagged isozyme PK1 in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
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 | Comment | Organism | Structure |
---|---|---|---|
ATP | inhibition of full-length enzyme at concentration above 2.5 mM | Staphylococcus aureus | |
D-fructose 1,6-bisphosphate | inhibition of full-length enzyme at 10 mM | Staphylococcus aureus | |
phosphate | - |
Staphylococcus aureus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Staphylococcus aureus | |
Mg2+ | required | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
53100 | - |
4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation | Staphylococcus aureus |
65100 | - |
4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation | Staphylococcus aureus |
215000 | - |
His-tagged C-terminally truncated enzyme mutant, gel filtration | Staphylococcus aureus |
250000 | - |
His-tagged full-length enzyme, gel filtration | Staphylococcus aureus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + pyruvate | Staphylococcus aureus | - |
ADP + phosphoenolpyruvate | - |
? | |
ATP + pyruvate | Escherichia coli | - |
ADP + phosphoenolpyruvate | - |
? | |
ATP + pyruvate | Staphylococcus aureus MRSA252 | - |
ADP + phosphoenolpyruvate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | A0A0H3JER5 | isozyme PK2; gene pykA | - |
Escherichia coli | P0AD62 | isozyme PK1; gene pykF | - |
Staphylococcus aureus | Q6GG09 | a methicillin-resistant strain, gene pyk | - |
Staphylococcus aureus MRSA252 | Q6GG09 | a methicillin-resistant strain, gene pyk | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged full-length and C-terminally truncated enzymes from Escherichia coli strain BL-21(DE3) by nickel affiniy chromatography | Staphylococcus aureus |
recombinant His-tagged isozyme PK1 from Escherichia coli strain BL21(DE3) by nickel affiniy chromatography | Escherichia coli |
recombinant His-tagged isozyme PK2 from Escherichia coli strain BL21(DE3) by nickel affiniy chromatography | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + pyruvate | - |
Staphylococcus aureus | ADP + phosphoenolpyruvate | - |
? | |
ATP + pyruvate | - |
Escherichia coli | ADP + phosphoenolpyruvate | - |
? | |
ATP + pyruvate | - |
Staphylococcus aureus MRSA252 | ADP + phosphoenolpyruvate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | - |
Escherichia coli |
homotetramer | 4 * 65100, about, full-length enzyme, sequence calculation, 4 * 53100, about, C-terminally truncated enzyme mutant, sequence calculation | Staphylococcus aureus |
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 | Comment | Organism |
---|---|---|
MRSA PK | - |
Staphylococcus aureus |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Staphylococcus aureus |
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Staphylococcus aureus |
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Staphylococcus aureus | |
ATP | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | significant evolutionary distance existing between the type I and type II isoenzymes in Gram-negative bacteria | Staphylococcus aureus |
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 |
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 |
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 |