Information on EC 2.7.1.68 - 1-phosphatidylinositol-4-phosphate 5-kinase

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The expected taxonomic range for this enzyme is: Eukaryota

EC NUMBER
COMMENTARY hide
2.7.1.68
-
RECOMMENDED NAME
GeneOntology No.
1-phosphatidylinositol-4-phosphate 5-kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate = ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
Phosphorylation
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
3-phosphoinositide biosynthesis
-
-
D-myo-inositol (1,4,5)-trisphosphate biosynthesis
-
-
Inositol phosphate metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:1-phosphatidyl-1D-myo-inositol-4-phosphate 5-phosphotransferase
This enzyme can also phosphorylate PtdIns3P in the 4-position, and PtdIns, PtdIns3P and PtdIns(3,4)P2 in the 5-position in vitro, but to a lesser extent. The last of these reactions occurs in vivo and is physiologically relevant. Three different isoforms are known.
CAS REGISTRY NUMBER
COMMENTARY hide
104645-76-3
-
9032-61-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
Eimeria maxima Weybridge strain
-
UniProt
Manually annotated by BRENDA team
-
TrEMBL
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + 1,2-dipalmitoyl phosphatidyl-1D-myo-inositol 4,5-bisphosphate
ADP + 1,2-dipalmitoyl phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
-
-
-
-
?
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-phosphatidyl-1D-myo-inositol-4,5-bisphosphate
show the reaction diagram
-
PIP5K3 is involved in localizing 1-phosphatidyl-1D-myo-inositol-4,5-bisphosphate to the elongating root hair apex and is a key regulator of the machinery that initiates and promotes root hair tip growth.
-
r
ATP + 1-stearoyl-2-arachidonoyl phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-stearoyl-2-arachidonoyl phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
-
-
-
-
?
ATP + 1-stearoyl-2-oleoyl phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-stearoyl-2-oleoyl phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
-
-
-
-
?
ATP + dipalmitoyl phosphatidylinositol 4-phosphate
ADP + dipalmitoylphosphatidylinositol 4,5-bisphosphate
show the reaction diagram
-
-
-
-
?
ATP + phosphatidylinositol 3-phosphate
ADP + phosphatidylinositol 3,4-bisphosphate
show the reaction diagram
-
-
-
?
ATP + phosphatidylinsositol 3,4-bisphosphate
ADP + phosphatidylinositol 3,4,5-trisphosphate
show the reaction diagram
-
-
-
-
?
GTP + 1-phosphatidyl-1D-myo-inositol 4-phosphate
GDP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate
show the reaction diagram
ATP + 1-phosphatidyl-1D-myo-inositol 4-phosphate
ADP + 1-phosphatidyl-1D-myo-inositol-4,5-bisphosphate
show the reaction diagram
O48709
-
PIP5K3 is involved in localizing 1-phosphatidyl-1D-myo-inositol-4,5-bisphosphate to the elongating root hair apex and is a key regulator of the machinery that initiates and promotes root hair tip growth.
-
r
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Ca2+
-
activates to a smaller extent than Mg2+
Fe2+
-
2 mM, slight inhibition
K+
70 mM in the activity assay
KCl
-
at 80 mM
MgCl2
-
at 10 mM
Zn2+
-
2 mM, slight inhibition
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate
2'(3')-O-(2,4,6-Trinitrophenyl)ATP
-
competitive
ATP
-
above 4 mM
Cetyltrimethylammonium bromide
-
-
Cutsum
-
-
-
cysteine
-
1 mM, 49% inhibition
EDTA
-
slight inhibition between 0.5-5 mM
GSSG
-
0.1 mM, 17% inhibition
heparin
ISP-1/myriocin
-
potent inhibitor, 750ng/ml
NEM
-
1 mM, 26% inhibition
PCMB
-
0.1 mM, complete inhibition
phosphatidic acid
phosphatidylinositol
-
0.1 mM, 125% stimulation
phosphatidylserine
-
0.1 mM, 125% stimulation
quercetin
small G proteins ARF
-
ARF activation absolutely requires the presence of phosphatidic acid, which cannot be substituted by phosphatidylserine, phosphatidylethanolamine, or lysophophatidic acid, but by competitive phosphatidylcholine, proteins ARF1 and ARF3, from brain cytosol, purification and identification, the 21 kDa G-proteins are activated by GTPgammaS, not by GDP, and activate themselves the PI(4)P 5-kinase
-
Sodium deoxycholate
-
-
spermine
-
0.03 mg/ml, 50% inhibition
Trifluoperazine
-
IC50: about 0.015 mM
Triton X-100
Wortmannin
-
phosphatidylinositol 4-kinase activity of STT4, but not of PIK1 is potently inhibited
ZnSO4
-
sensitive to 9 mM ZnSO4
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,2-diarachidonoyl phosphatidic acid
-
-
1,2-dilinoleoyl phosphatidic acid
-
-
1,2-dioleoyl phosphatidic acid
-
-
1-arachidoyl-2-arachidonoyl phosphatidic acid
-
-
1-stearoyl-2-arachidonoyl phosphatidic acid
-
-
1-stearoyl-2-oleoyl phosphatidic acid
-
-
Acetylcholine
-
0.01 mM or 0.1 mM, each in presence of 0.1 mM eserine, stimulates the enzyme in homogenates by 30-40%
ADP-ribosylation factor 1
-
0.001 mM
-
ADP-ribosylation factor 6
-
0.001 mM
-
advanced glycation end products
Ajuba
-
dramatic activation of enzymatic activity
-
AP-2mu-endocytic cargo complex
-
clathrin adaptor complex, leads to a potent stimulation of PIPK activity
-
Arf6
-
activates the enzyme via binding of small GTPase Rac1, regulatory function, mutant Arf6 N121I is defective for nucleotide binding, mutant Q67L is similarly active as the wild-type Arf6
-
beta2 adaptin
-
the beta2 subunit of beta2 adaptin binds to the C-terminal tail of PIP5Kgamma661 and causes activation of the enzyme
-
heparin
-
type I enzyme is stimulated at low concentrations
lipid phosphatidate
-
0.1 mM
-
lysophosphatidic acid
myelin basic protein
-
enhances activity when phosphatidylinositol 4-phosphate/phosphatidylethanolamine vesicles are used as substrate
-
phosphatidic acid
small G protein ARF
-
ARF activation absolutely requires the presence of phosphatidic acid, which cannot be substituted by phosphatidylserine, phosphatidylethanolamine, or lysophosphatidic acid, but by competitive phosphatidylcholine, reconstitution of the recombinant either isozyme alpha, beta, or gamma with recombinant class II and III ARFs, ARF5 and ARF6, respectively, as well as myristoylated class I ARF in presence of GTPgammaS or GDP, and phosphatidic acid activates the isozymes
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small G protein RHO
-
activates the enzyme in neurits, crucial in cell shapig together with the enzyme, overview
-
small G proteins
-
-
-
small G proteins ARF
small GTPase Arf
small GTPase ARF/phosphatidic acid
-
regulation of isozyme gamma
-
small GTPase Rho
spermidine
-
enhances activity
spermine
Triton X-100
additional information
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.262
1-phosphatidyl-1D-inositol 4-phosphate
-
pH 7.5, enzyme type Ibeta
0.005 - 0.065
1-phosphatidyl-1D-myo-inositol 3-phosphate
0.0012 - 4.1
1-phosphatidyl-1D-myo-inositol 4-phosphate
4.9 - 16
1-stearoyl-2-arachidonoyl phosphatidyl-1D-myo-inositol 4-phosphate
1.6 - 3.7
1-stearoyl-2-oleoyl phosphatidyl-1D-myo-inositol 4-phosphate
0.002 - 0.2
ATP
0.133
GTP
-
pH 7.5, 22C
0.129 - 0.13
MgATP2-
0.006 - 0.08
phosphatidylinositol 3,4-bisphosphate
0.12
phosphatidylinositol 3-phosphate
-
pH 7.5, enzyme type IIalpha
additional information
additional information
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.055
2'(3')-O-(2,4,6-Trinitrophenyl)ATP
-
pH 7.4, 30C
0.035
quercetin
-
pH 7.5, 25C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0001
Ca2+
Bos taurus
-
free Ca2+, IC50: about 0.0001 mM
0.015
Trifluoperazine
Bos taurus
-
IC50: about 0.015 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.015
-
membrane bound type II enzyme
0.023
-
cytosolic enzyme
0.085
-
-
0.117
-
-
0.171
-
-
1.75
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5 - 7
-
shoot and root plasma membrane, endogenous substrate
7.2
-
-
7.4
-
assay at
7.5 - 8.2
-
shoot plasma membrane, exogenous substrate
7.8 - 8.2
-
root plasma membrane, exogenous substrate
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 8
-
pH 5.0: about 65% of maximal activity, pH 8.0: about 75% of maximal activity
5.5 - 8.5
-
pH 5.5: about 35% of maximal activity, pH 8.5: about 60% of maximal activity
6.1 - 7.9
-
pH 6.1: about 85% of maximal activity, pH 7.9: about 85% of maximal activity
6.5 - 8.5
-
pH 6.5: about 35% of maximal activity, pH 8.5: about 60% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.8
-
isoelectric focusing
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
isozymes type I PIP5Kalpha, PIP5Kbeta, and PIP5Kgamma are all expressed in BV-2 microglial cells
Manually annotated by BRENDA team
Escherichia coli BL21 cell is used for cloning and expression
Manually annotated by BRENDA team
-
granule cells
Manually annotated by BRENDA team
-
high expression of PIP5KIbeta and PIP5KIalpha
Manually annotated by BRENDA team
enzyme form 10, most abundant in
Manually annotated by BRENDA team
-
low expression of PIP5KIalpha
Manually annotated by BRENDA team
-
enzyme is enriched in the uropod during chemotaxis of primary neutrophils. Enrichment occurs early upon chemoattractant stimulation and is persistent during chemotaxis, concomitant with enrichment of phosphatidylinositol 4,5-bisphosphate
Manually annotated by BRENDA team
-
moderate expression of PIP5KIbeta, high expression of PIP5KIalpha
Manually annotated by BRENDA team
-
high expression of PIP5KIbeta
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
localizes mainly to early endosomes. PIKfyve is distributed in microdomains
Manually annotated by BRENDA team
-
co-localization of KIF2A and PIPKalpha at the tips of neurites
-
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
87000
-
SDS-PAGE
130000
-
gel filtration
150000
-
gel filtration
237000
calculated from sequence
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
-
x * 60000, SDS-PAGE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
sucrose and PEG 20000 in the assay medium are required to stabilize the enzyme during phosphorylation reaction
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 50 MM Tris-HCl, pH 8.0, 1 mM EDTA, 0.25 M sucrose, 0.3% v/v 2-mercaptoethanol, 0.1% Triton X-100, 0.1% PEG 20000, 0.05 mM ATP, 0.1 mM PMSF, stable for several months
-
-70C, stable in concentrated form
-
-80C, stable for at least 3 months
-
4C, about 10% loss of activity after overnight storage
-
4C, storage causes the 53000 Da enzyme to be slowly degraded to a 45000 Da peptide
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
cytosolic enzyme type II enzyme and membrane-bound type I and type II enzyme
-
glutathione-Sepharose chromatography
-
Partial purification from Escherichia coli culture cells. The cells are suspended in PBS buffer, sonicated on ice, and centrifuged. Supernatant is mixed with glutathione-Sepharose 4B and incubated at 48C for 2 h. The Sepharose beads are collected and washed with an excess volume of PBS.
recombinan GST-fusion isozyme alpha from Escherichia coli
recombinant GST-fusion protein from Escherichia coli in presence of Triton X-100 and PtdOH; recombinant GST-fusion protein from Escherichia coli in presence of Triton X-100 and PtdOH
Talon metal affinity column chromatography
-
wild-type and mutant isozyme gamma, partially by immunoprecipitation and protein G affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a yellow fluorescent (YFP) und GUS fusion PIP5K3 protein is expressed in Escherichia coli cells and transformed in Arabidopsis thaliana to produce trangenic plants. Overexpression and mutant is investigated.
cloning of splice variant phosphatidylinositol 4-phosphate 5-kinase Igamma_v6 from hippocampus, expression in COS-7 cells
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cloning of splice variant phosphatidylinositol 4-phosphate 5-kinase Igamma_v6, expression in COS-7 cells
-
cloning of splice variant phosphatidylinositol 4-phosphate 5-kinase Igamma_v6, expression in COS-7 cells. In humans there is an alternative splice site 78 residues into exon 16c that splices out the rest of the Igamma_v5 insert, and then jumps either 1. to the start of exon 17 to generate human Igamma_v3 (the stop codon is then at the start of human exon 18), or 2. straight to that stop codon in exon 18 to generate human Igamma_v6, cloning of splice variants phosphatidylinositol 4-phosphate 5-kinase Igamma_v3, and Igamma_i2 and expression analysis
-
coexpression of wild-type HA-tagged isozyme alpha or HA-tagged isozyme alpha mutant D227A with HA-tagged Arf6 or HA-tagged Arf6 mutant N121I in COS-1 cells
-
EmPIP5K gene, DNA and amino acid sequence determination and analysis, located at position 710-8036 nt on the complimentary strand, structural organization of the PIP5K gene, and phylogenetic analysis and tree
enzyme form PIP5KIalpha
-
expressed in Escherichia coli
-
expressed in Escherichia coli strain HT115 (DE3)
-
expressed in Escherichia coli; expressed in HEK-293T cells
-
expressed in HEK-293T cells; expression in COS-7 cells
-
expressed in HeLa cells
-
expressed in Nicotiana tabacum
expressed in Sf9 cells and MEF cells
-
expression in COS cells
-
expression in Escherichia coli
-
expression in HEK-203 cell
-
expression in Xenopus laevis oocyte
-
expression of enzyme form 10 as GST-fusion protein in Escherichia coli, expression of His-tagged enzyme form 10 in Spodoptera frugiperda Sf9 cells via baculovirus infection system; expression of enzyme form 1 as GST-fusion protein in Escherichia coli, expression of His-tagged enzyme form 1 in Spodoptera frugiperda Sf9 cells via baculovirus infection system
expression of HA-tagged PIP5K isozymes in COS-7 cells, expression of isozyme PIP5Kalpha mutant D322A
-
expression of isozyme alpha as GST-fusion protein in Escherichia coli
expression of isozyme PIP5Kalpha tagged with monomeric red fluorescent protein, haemagglutinin, or FLAG in HEK-293T cells
expression of mutant enzymes K181A, D309N, D309N/R429Q and R427Q in Escherichia coli, expression of mutant enzymes in human MG-63 osteosarcoma cells
-
functional complementation of a yeast Mss4-1 mutant strain YOC808 by the wild-type isozyme gamma and the deletion mutants localizing to the plasma membrane, while the deletion mutants localized to the cytosol cannot support yeast cell growth, expression of wild-type and mutant type I HA-tagged or EGFP-tagged isozymes gamma in Neuro 2A cells and in COS-1 cells
-
functional expression in HeLa cells as GFP-fusion protein, translocation to the plasma membrane by coexpression of ARF6 upon stimulation by EGF, coexpression of ARF6 mutant N122I inhibits the membrane ruffling, overview
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identification of splicing form c of type I phosphatidylinositol 4-phosphate 5-kinase isoform gamma possessing additional 26 amino acids at the C-terminus from a hippocampus library, DNA and amino acid sequence determination and analysis, overvexpression of the catalytically inactive mutants D316K and K188A of isozyme gamma splicing form c in rat cerebellar granule cells, expression of isozyme gamma splicing form a, expression of wild-type and FLAG-tagged isozyme gamma splicing form c in COS-7 cells
-
isozyme PIP5K1beta expression in HEK-293T and COS-7 cells
-
overexpression of isozymes alpha, beta, and gamma in HeLa cells, overexpression of isozymes alpha, beta, and gamma in CV-1 cells: isozymes alpha and beta are active, the latter shows the highest activity, isozyme gamma is not active, small interference RNA inhibition of isozyme beta expression also reduces expression of isozyme alpha, but increased expression of isozyme gamma, with a 50% loss in activity, reduction of isozyme alpha expression increases expression of isozymes beta and gamma and leaves the 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate synthesis level unaltered, inhibition of isozyme gamma increases expression of isozymes alpha and beta, overview, expression of isozyme beta increases the internalization of influenza virus hemagglutinin Y543, enzyme expression prevents the cells of disruption of their actine cytoskeleton by drugs, e.g. cytochalasin D or latrunculin, and increases the number of clathrin-coated pits
-
overexpression of mutant D266A in N1E-115 cells, expression of isozyme alpha in COS-7 cells, expression of isozyme alpha in Escherichia coli
-
overexpression of PIPKalpha in N1E-115 neuroblastoma cells, co-expression of His6-tagged KIF2A and GST-tagged PIPKalpha in Escherichia coli, binding analysis
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D309N
-
inactive mutant
R427Q
-
inactive mutant
D203A
-
catalytically inactive. Expression in Xenopus laevis oocyte has no effect on epithelial sodium channel activity
D253A
-
mutant lacking kinase activity. Overexpression compromises uropod formation and rear retraction during chemotaxis
D266A
-
site-directed mtagenesis, mutation of the putative ATP and 1-phosphatidyl-1D-myo-inositol 4-phosphate binding site, cells form extremely extended bipolar neurits even in presence of serum, failed to interfere with cell flattening and neurite extension
D309N
-
greatly reduced kinase activity towards 1-phosphatidyl-1D-myo-inositol 4-phosphate
D309N/R427Q
-
greatly reduced kinase activity towards 1-phosphatidyl-1D-myo-inositol 4-phosphate
D316K
-
no activity
D322A
-
isozyme PIP5Kalpha mutant that shows reduced activity compared to the wild-type enzyme
K138A
-
no activity
K181A
-
greatly reduced kinase activity towards 1-phosphatidyl-1D-myo-inositol 4-phosphate
L202I
-
the mutations of PIP5Kalpha decreases the substrate affinity and the enzyme efficiency for 1-stearoyl-2-arachidonoyl phosphatidylinositol-4-phosphate compared to the wild-type enzyme
L210I
-
the mutations of PIP5Kalpha decreases the substrate affinity and the enzyme efficiency for 1-stearoyl-2-arachidonoyl phosphatidylinositol-4-phosphate compared to the wild-type enzyme
R332H
YFP-tagged tubby mutant
R427Q
-
greatly reduced kinase activity towards 1-phosphatidyl-1D-myo-inositol 4-phosphate
D316K
-
site-directed mutagenesis, overexpression of the catalytically inactive construct of isozyme gamma splicing form c in rat cerebellar granule cells causes progressive loss of their neuronal processes, not observed with expression of isozyme gamma splicing form a
K188A
-
site-directed mutagenesis, overexpression of the catalytically inactive construct of isozyme gamma splicing form c in rat cerebellar granule cells causes progressive loss of their neuronal processes, not observed with expression of isozyme gamma splicing form a
D598A
-
no activity
D636A
-
no activity
additional information
Show AA Sequence (227 entries)
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