Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.7.1.40 extracted from

  • Wulfert, S.; Schilasky, S.; Krueger, S.
    Transcriptional and biochemical characterization of cytosolic pyruvate kinases in Arabidopsis thaliana (2020), Plants (Basel), 9, 353 .
    View publication on PubMed
Search for more literature for 2.7.1.40

Activating Compound

Activating Compound Comment Organism Structure
AMP 9% activation at 0.1 mM, cPK3 Arabidopsis thaliana
fructose-1,6-bisphosphate FBP, 22% activation at 1 mM, cPK1 Arabidopsis thaliana
fructose-1,6-bisphosphate FBP, 4% activation at 1 mM, cPK2 Arabidopsis thaliana
L-serine 12% activation at 0.2 mM, cPK2 Arabidopsis thaliana
additional information no effect by L-serine and L-glutamate at 0.2 mM on cPK4, poor effect by fructose-1,6-bisphosphate at 1.0 mM Arabidopsis thaliana
additional information no effect by L-serine and L-glutamate at 0.2 mM on cPK5 Arabidopsis thaliana
additional information poor effect by AMP at 0.1 mM on cPK1 Arabidopsis thaliana
additional information poor effect by fructose-1,6-bisphosphate at 1 mM and L-glutamate at 0.2 mM on cPK3 Arabidopsis thaliana

Cloned(Commentary)

Cloned (Comment) Organism
gene at2g36580, phylogenetic tree of Arabidopsis thaliana isozymes, recombinant expression of YFP-tagged isozymes in in Nicotiana benthamiana leaf cells via Agrobacterium tumefaciens-mediated transformation, recombinant expression of N-terminally His5-tagged enzyme in Escherichia coli Arabidopsis thaliana
gene at3g52990, phylogenetic tree of Arabidopsis thaliana isozymes, recombinant expression of YFP-tagged isozymes in in Nicotiana benthamiana leaf cells via Agrobacterium tumefaciens-mediated transformation, recombinant expression of N-terminally His5-tagged enzyme in Escherichia coli Arabidopsis thaliana
gene at5g08570, phylogenetic tree of Arabidopsis thaliana isozymes, recombinant expression of YFP-tagged isozymes in in Nicotiana benthamiana leaf cells via Agrobacterium tumefaciens-mediated transformation, recombinant expression of N-terminally His5-tagged enzyme in Escherichia coli Arabidopsis thaliana
gene at5g56350, phylogenetic tree of Arabidopsis thaliana isozymes, recombinant expression of YFP-tagged isozymes in in Nicotiana benthamiana leaf cells via Agrobacterium tumefaciens-mediated transformation, recombinant expression of N-terminally His5-tagged enzyme in Escherichia coli Arabidopsis thaliana
gene at5g63680, phylogenetic tree of Arabidopsis thaliana isozymes, recombinant expression of YFP-tagged isozymes in in Nicotiana benthamiana leaf cells via Agrobacterium tumefaciens-mediated transformation, recombinant expression of N-terminally His5-tagged enzyme in Escherichia coli Arabidopsis thaliana

Inhibitors

Inhibitors Comment Organism Structure
AMP 90% inhibition at 0.1 mM, cPK2 Arabidopsis thaliana
ATP 16% inhibition at 2 mM, cPK4; 24% inhibition at 2 mM, cPK2; 5% inhibition at 2 mM, cPK5; 73% inhibition at 2 mM, cPK3; 93% inhibition at 2 mM, cPK1 Arabidopsis thaliana
citrate 74% inhibition at 4 mM, cPK2; 83% inhibition at 4 mM, cPK1; 93% inhibition at 4 mM, cPK3; 97% inhibition at 4 mM, cPK4; 98% inhibition at 4 mM, cPK5 Arabidopsis thaliana
L-aspartate 31% inhibition at 0.2 mM, cPK1; 6% inhibition at 0.2 mM, cPK3 Arabidopsis thaliana
L-glutamate 15% inhibition at 0.2 mM, cPK1 Arabidopsis thaliana
additional information no effect by L-serine and L-glutamate at 0.2 mM on cPK4, poor effects by L-aspartate at 0.2 mM and AMP at 0.1 mM; no effect by L-serine and L-glutamate at 0.2 mM on cPK5, poor effects by L-aspartate at 0.2 mM and AMP at 0.1 mM, and by fructose-1,6-bisphosphate at 1.0 mM; poor effect by L-serine at 0.2 mM on cPK3; poor effect by serine at 0.2 mM on cPK1; poor effects by L-glutamate and L-aspartate at 0.2 mM on cPK2 Arabidopsis thaliana

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information Michaelis-Menten kinetics Arabidopsis thaliana
0.03
-
 ADP cPK2, pH 7.5, 25°C Arabidopsis thaliana
0.05
-
 phosphoenolpyruvate cPK5, pH 7.5, 25°C Arabidopsis thaliana
0.06
-
 phosphoenolpyruvate cPK1, pH 7.5, 25°C Arabidopsis thaliana
0.07
-
 ADP cPK4, pH 7.5, 25°C Arabidopsis thaliana
0.14
-
 ADP cPK3, pH 7.5, 25°C Arabidopsis thaliana
0.15
-
 ADP cPK1, pH 7.5, 25°C Arabidopsis thaliana
0.17
-
 phosphoenolpyruvate cPK3, pH 7.5, 25°C Arabidopsis thaliana
0.17
-
 phosphoenolpyruvate cPK4, pH 7.5, 25°C Arabidopsis thaliana
0.34
-
 ADP cPK5, pH 7.5, 25°C Arabidopsis thaliana
0.76
-
 phosphoenolpyruvate cPK2, pH 7.5, 25°C Arabidopsis thaliana

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol determination of five cytosol-localized pyruvate kinases, out of the fourteen putative pyruvate kinase genes encoded by the Arabidopsis thaliana genome, expression analysis, overview Arabidopsis thaliana 5829
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Arabidopsis thaliana

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + phosphoenolpyruvate Arabidopsis thaliana
-
 ATP + pyruvate
-
 ?

Organism

Organism UniProt Comment Textmining
Arabidopsis thaliana Q94KE3
-
-
Arabidopsis thaliana Q9FFP6
-
-
Arabidopsis thaliana Q9FM97
-
-
Arabidopsis thaliana Q9FNN1
-
-
Arabidopsis thaliana Q9SJQ0
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His5-tagged enzyme from Escherichia coli by nickel affinity chromatography and desalting gel filtration Arabidopsis thaliana

Source Tissue

Source Tissue Comment Organism Textmining
cotyledon
-
 Arabidopsis thaliana
-
leaf
-
 Arabidopsis thaliana
-
leaf vasculature and mesophyll of rosette leaves Arabidopsis thaliana
-
leaf young, vasculature and mesophyll of rosette leaves Arabidopsis thaliana
-
mesophyll cell
-
 Arabidopsis thaliana
-
additional information isozyme localization analysis revealing an isoform-specific expression pattern for the five cytosolic enzymes, subject to plant tissue and developmental stage. Cytosolic pyruvate kinase genes are expressed in a tissue-specific and developmental-specific manner. cPK1 promoter GUS lines display strong staining in the shoot apical meristem and in the basal part of young leaves. The cPK1 promoter leads to broadly abundant GUS activity in roots and leaves of seedlings from the first day on Arabidopsis thaliana
-
additional information isozyme localization analysis revealing an isoform-specific expression pattern for the five cytosolic enzymes, subject to plant tissue and developmental stage. Cytosolic pyruvate kinase genes are expressed in a tissue-specific and developmental-specific manner. cPK4-promoter plants expression is observed from the first day on in the entire root and the leaf vasculature, and later in hydathodes and at the sides of emerging leaf buds Arabidopsis thaliana
-
additional information isozyme localization analysis revealing an isoform-specific expression pattern for the five cytosolic enzymes, subject to plant tissue and developmental stage. Cytosolic pyruvate kinase genes are expressed in a tissue-specific and developmental-specific manner. Expression of cPK5 starts later and is mainly restricted to the cotyledon, leaf and root vasculature Arabidopsis thaliana
-
additional information isozyme localization analysis revealing an isoform-specific expression pattern for the five cytosolic enzymes, subject to plant tissue and developmental stage. Cytosolic pyruvate kinase genes are expressed in a tissue-specific and developmental-specific manner. The cPK2 promoter shows broadly abundant GUS signals only at later stages, cPK2 promoter GUS expression does not initiate in the rosette axis until ten days after germination Arabidopsis thaliana
-
additional information isozyme localization analysis revealing an isoform-specific expression pattern for the five cytosolic enzymes, subject to plant tissue and developmental stage. Cytosolic pyruvate kinase genes are expressed in a tissue-specific and developmental-specific manner. The cPK3 promoter shows broadly abundant GUS signals only at later stages. The cPK3 promoter-induced expression is restricted to the root tip and is already visible on the first day after germination. From the third day on, the signal becomes abundant in the entire root and is also observed in the leaf axis and spatial related trichomes Arabidopsis thaliana
-
root
-
 Arabidopsis thaliana
-
root root tip Arabidopsis thaliana
-
seedling
-
 Arabidopsis thaliana
-
shoot
-
 Arabidopsis thaliana
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + phosphoenolpyruvate
-
 Arabidopsis thaliana ATP + pyruvate
-
 ?

Synonyms

Synonyms Comment Organism
CPK
-
 Arabidopsis thaliana
CPK1
-
 Arabidopsis thaliana
cPK2
-
 Arabidopsis thaliana
cPK3
-
 Arabidopsis thaliana
cPK4
-
 Arabidopsis thaliana
cPK5
-
 Arabidopsis thaliana
cytosolic pyruvate kinase
-
 Arabidopsis thaliana

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Arabidopsis thaliana

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.53
-
 ADP cPK1, pH 7.5, 25°C Arabidopsis thaliana
1.53
-
 phosphoenolpyruvate cPK1, pH 7.5, 25°C Arabidopsis thaliana
2.4
-
 ADP cPK3, pH 7.5, 25°C Arabidopsis thaliana
2.4
-
 phosphoenolpyruvate cPK3, pH 7.5, 25°C Arabidopsis thaliana
3.05
-
 ADP cPK2, pH 7.5, 25°C Arabidopsis thaliana
3.05
-
 phosphoenolpyruvate cPK2, pH 7.5, 25°C Arabidopsis thaliana
4
-
 ADP cPK4, pH 7.5, 25°C Arabidopsis thaliana
4
-
 phosphoenolpyruvate cPK4, pH 7.5, 25°C Arabidopsis thaliana
6.68
-
 ADP cPK5, pH 7.5, 25°C Arabidopsis thaliana
6.68
-
 phosphoenolpyruvate cPK5, pH 7.5, 25°C Arabidopsis thaliana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Arabidopsis thaliana

Expression

Organism Comment Expression
Arabidopsis thaliana the cytosolic PKs are induced in response to cold treatment. cPK1 and cPK2 expression is induced, whereas cPK3 transcript levels are not altered. The strongest induction upon cold treatment is found for the isogenes cPK4 and cPK5 up

General Information

General Information Comment Organism
evolution 14 putative pyruvate kinase genes are encoded by the Arabidopsis thaliana genome. Determination of five cytosol-localized pyruvate kinases, out of the fourteen putative pyruvate kinase genes encoded by the Arabidopsis thaliana genome, phylogenetic analysis. The five identified cytosolic pyruvate kinase isoforms adjust the carbohydrate flux through the glycolytic pathway in Arabidopsis thaliana, by distinct regulatory qualities, such as individual expression pattern as well as dissimilar responsiveness to allosteric effectors and enzyme subgroup association Arabidopsis thaliana
additional information the isozymes cPK1-5 show positive, synergistic effects when mixed Arabidopsis thaliana
physiological function cytosolic PK is a key player in energy allocation, as it generates ATP in the cytosol, supplies pyruvate to the TCA cycle and thereby drives mitochondrial ATP synthesis. The cytosolic pyruvate kinase (cPK) represents a key glycolytic enzyme by catalyzing phosphate transfer from phosphoenolpyruvate (PEP) to ADP for the synthesis of ATP. Besides its important functions in cellular energy homeostasis, the activity of cytosolic pyruvate kinase underlies tight regulation, for instance by allosteric effectors, that impact stability of its quaternary structure. The five identified cytosolic pyruvate kinase isoforms adjust the carbohydrate flux through the glycolytic pathway in Arabidopsis thaliana, by distinct regulatory qualities, such as individual expression pattern as well as dissimilar responsiveness to allosteric effectors and enzyme subgroup association Arabidopsis thaliana
physiological function cytosolic PK is a key player in energy allocation, as it generates ATP in the cytosol, supplies pyruvate to the TCA cycle and thereby drives mitochondrial ATP synthesis. The cytosolic pyruvate kinase (cPK) represents a key glycolytic enzyme by catalyzing phosphate transfer from phosphoenolpyruvate (PEP) to ADP for the synthesis of ATP. Besides its important functions in cellular energy homeostasis, the activity of cytosolic pyruvate kinase underlies tight regulation, for instance by allosteric effectors, that impact stability of its quaternary structure. The five identified cytosolic pyruvate kinase isoforms adjust the carbohydrate flux through the glycolytic pathway in Arabidopsis thaliana, by distinct regulatory qualities, such as individual expression pattern as well as dissimilar responsiveness to allosteric effectors and enzyme subgroup association. Spatial distribution of glycolytic enzymes within the cell constitutes a further point of regulation, since enzymes may localize at sites of demand for glycolytic intermediates Arabidopsis thaliana

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.01
-
 phosphoenolpyruvate cPK2, pH 7.5, 25°C Arabidopsis thaliana
10.2
-
 ADP cPK1, pH 7.5, 25°C Arabidopsis thaliana
14.12
-
 phosphoenolpyruvate cPK3, pH 7.5, 25°C Arabidopsis thaliana
17.14
-
 ADP cPK3, pH 7.5, 25°C Arabidopsis thaliana
19.65
-
 ADP cPK5, pH 7.5, 25°C Arabidopsis thaliana
23.53
-
 phosphoenolpyruvate cPK4, pH 7.5, 25°C Arabidopsis thaliana
25.5
-
 phosphoenolpyruvate cPK1, pH 7.5, 25°C Arabidopsis thaliana
57.14
-
 ADP cPK4, pH 7.5, 25°C Arabidopsis thaliana
101.67
-
 ADP cPK2, pH 7.5, 25°C Arabidopsis thaliana
133.6
-
 phosphoenolpyruvate cPK5, pH 7.5, 25°C Arabidopsis thaliana