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Information on EC 2.7.1.32 - choline kinase and Organism(s) Plasmodium falciparum

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IUBMB Comments
Ethanolamine and its methyl and ethyl derivatives can also act as acceptors.
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This record set is specific for:
Plasmodium falciparum
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The taxonomic range for the selected organisms is: Plasmodium falciparum
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
choline kinase, choline kinase alpha, chokalpha, choline kinase beta, choline/ethanolamine kinase, chk-alpha, chokalpha1, ck-alpha, schok, choline kinase-alpha, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ATP:choline phosphotransferase
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CHETK-alpha
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-
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ChoK
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-
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choline kinase
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choline phosphokinase
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choline-ethanolamine kinase
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choline/ethanolamine kinase
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EtnK
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kinase, choline (phosphorylating)
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
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-
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Phosphorylation
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-
PATHWAY SOURCE
PATHWAYS
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-, -, -, -
SYSTEMATIC NAME
IUBMB Comments
ATP:choline phosphotransferase
Ethanolamine and its methyl and ethyl derivatives can also act as acceptors.
CAS REGISTRY NUMBER
COMMENTARY hide
9026-67-9
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SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + choline
ADP + O-phosphocholine
show the reaction diagram
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-
-
-
?
ATP + choline
ADP + phosphocholine
show the reaction diagram
ATP + ethanolamine
ADP + ethanolamine phosphate
show the reaction diagram
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can not phosphorylate ethanolamine efficiently
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-
?
choline + ATP
phosphocholine + ADP
show the reaction diagram
-
choline kinase may have a regulatory role in phosphatidylcholine biosynthesis
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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
ATP + choline
ADP + phosphocholine
show the reaction diagram
-
-
-
?
choline + ATP
phosphocholine + ADP
show the reaction diagram
-
choline kinase may have a regulatory role in phosphatidylcholine biosynthesis
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-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mg2+
required, enzyme binding structure analysis. The ADP molecule also participates in the coordination of two magnesium ions
MgCl2
-
assay at
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ethanolamine
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poor inhibitor
H-89
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no activity on choline kinase at 5 mM ATP, decreasing the ATP concentration to 100 microM has no effect on choline kinase
HC-3
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poor inhibition of 16% and 18% is observed at 50 microM and 150 microM, respectively, at both 100 and 250 microM choline
hemicholinium-3
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poor inhibition of 16% and 18% at 0.050 mM and 0.15 mM, respectively, not significantly inhibited by hemicholinium-3 up to 0.025 mM
hexadecyltrimethylammonium bromide
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60% inhibition at 0.1 mM; HDTAB, structural resemblance to hexadecylphosphocholine, exhibits an antimalarial effect and inhibits choline kinase in a dose-dependent manner. For purified protein: 60% inhibition at 100 microM (in presence of 250 microM choline), 60% inhibition at 50 microM (in presence of 100 microM choline). HDTAB may compete with choline for the choline binding site of choline kinase and may offer competitive inhibition with respect to choline. The concentration of HDTAB required to inhibit Plasmodium falciparum growth by 62% was 10times lower than the concentration required to inhibit purified choline kinase by 60% in vitro
phosphocholine
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40% and 66% inhibition at 2.5 mM and 10 mM, respectively
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.5
ATP
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in 100 mM Tris/HCl, 6 mM MgCl2, 5 mM ATP, pH 8.8, at 37°C
0.145
choline
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in 100 mM Tris/HCl, 6 mM MgCl2, 5 mM ATP, pH 8.8, at 37°C
5
ethanolamine
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in 100 mM Tris/HCl, 6 mM MgCl2, 5 mM ATP, pH 8.8, at 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5.5
ethanolamine
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in 100 mM Tris/HCl, 6 mM MgCl2, 5 mM ATP, pH 8.8, at 37°C
3.45
phosphocholine
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in 100 mM Tris/HCl, 6 mM MgCl2, 5 mM ATP, pH 8.8, at 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
parasites isolated from infected human erythrocytes
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
selective inhibition of the enzyme and the Kennedy pathway leads to the parasite arrest and eradication
metabolism
within the Kennedy pathway, the Plasmodium falciparum choline kinase (PfChoK) has a central role in the biosynthesis of phosphatidylcholine. Plasmodium falciparum plasma membrane is mainly composed by phosphatidylcholine (PC) and phosphatidylethanolamine (PE), which represent 40-50% and 35-40% of the total phospholipid content, respectively
additional information
substrate binding structure analysis, overview
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52200
-
calculated from amino acid sequence
56000
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SDS-PAGE, recombinant protein
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinat His6-tagged choline kinase in apoform and in ADP-bound form, hanging drop vapor diffusion method, mixing of 0.001 ml of 10 mg/ml protein solution with 0.001 ml of reservoir solution, and equilibration against 0.5 ml reservoir solution, containing 16% v/v PEG 8000, 0.2 M NaCl, 0.1 M HEPES, pH 7.5, 2 mM Tris(2-carboxyethyl)phosphine (TCEP), and 4 mM MgCl2 for the apoform crystals, and the same composition with 2 mM ADP for the complex crystals, at room temperature, X-ray diffraction structrue determination and analysis at 2.0-2.2 A resolution, molecular replacement and model building using the PDB ID 3FI8 structure as starting model
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
Ni-nitrilotriacetic acid agarose affinity and gel filtration
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Ni-NTA agarose resin column chromatography and 300SW gel filtration
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overexpressed PfCK protein is purified to homogeneity using Ni-nitrilotriacetic acid agarose affinity and gel filtration chromatography
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recombinant N-terminally His6-tagged enzyme from Escherichia coli strain BL21(DE3) Star by nickel affinity chromatography and gel filtration
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli
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recombinant expression of N-terminally His6-tagged enzyme in Escherichia coli strain BL21(DE3) Star
The RT-PCR-amplified PfCK gene is cloned in pRSET-C, an Escherichia coli expression vector. The host strain is optimized
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
targeting the parasite lipid metabolism and choline kinase for pharmacological action constitutes an effective way of dealing with the spreading of the disease and with conventional therapy-resistant strains
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Choubey, V.; Maity, P.; Guha, M.; Kumar, S.; Srivastava, K.; Puri, S.K.; Bandyopadhyay, U.
Inhibition of Plasmodium falciparum choline kinase by hexadecyltrimethylammonium bromide: a possible antimalarial mechanism
Antimicrob. Agents Chemother.
51
696-706
2007
Plasmodium falciparum
Manually annotated by BRENDA team
Choubey, V.; Guha, M.; Maity, P.; Kumar, S.; Raghunandan, R.; Maulik, P.R.; Mitra, K.; Halder, U.C.; Bandyopadhyay, U.
Molecular characterization and localization of Plasmodium falciparum choline kinase
Biochim. Biophys. Acta
1760
1027-1038
2006
Plasmodium falciparum
Manually annotated by BRENDA team
Torretta, A.; Lopez-Cara, L.; Parisini, E.
Crystal structure of the apo and the adp-bound form of choline kinase from Plasmodium falciparum
Crystals
10
1-13
2020
Plasmodium falciparum (Q8IM71)
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Manually annotated by BRENDA team