Information on EC 3.1.3.36 - phosphoinositide 5-phosphatase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
3.1.3.36
-
RECOMMENDED NAME
GeneOntology No.
phosphoinositide 5-phosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O = 1-phosphatidyl-1D-myo-inositol 4-phosphate + phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
3-phosphoinositide degradation
-
-
Inositol phosphate metabolism
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphatidyl-myo-inositol-4,5-bisphosphate 4-phosphohydrolase
These enzymes can also remove the 5-phosphate from Ins(1,4,5)P3 and/or Ins(1,3,4,5)P4. They are a diverse family of enzymes, with differing abilities to catalyse two or more of the four reactions listed. They are thought to use inositol lipids rather than inositol phosphates as substrates in vivo. All of them can use either or both of PtdIns(4,5)P2 and PtdIns(3,4,5)P3 as substrates; this is the main property that distinguishes them from EC 3.1.3.56, inositol-polyphosphate 5-phosphatase.
CAS REGISTRY NUMBER
COMMENTARY hide
9036-01-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ecotype Col-0
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
and isogenic derivatives
-
-
Manually annotated by BRENDA team
and isogenic derivatives
-
-
Manually annotated by BRENDA team
serovar typhimurium
-
-
Manually annotated by BRENDA team
a Ca2+-dependent enzyme and a Mg2+-dependent enzyme
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
1-phosphatidyl-1D-myo-inositol 3,4,5-triphosphate + H2O
1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate + phosphate
show the reaction diagram
1-phosphatidyl-1D-myo-inositol 3,5-bisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 3-phosphate + phosphate
show the reaction diagram
-
-
-
-
?
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4-phosphate + phosphate
show the reaction diagram
1D-myo-inositol 1,2,4,5,6-pentakisphosphate + H2O
1D-myo-inositol 1,2,4,6-tetrakisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1D-myo-inositol 1,2,4,5-tetrakisphosphate + H2O
1D-myo-inositol 1,2,4-trisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1D-myo-inositol 1,3,4,5-tetrakisphosphate + H2O
1D-myo-inositol 1,3,4-trisphosphate + phosphate
show the reaction diagram
1D-myo-inositol 1,3,4,5-tetrasphosphate + H2O
1D-myo-inositol 1,3,4-trisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1D-myo-inositol 1,4,5-trisphosphate + H2O
1D-myo-inositol 1,4-bisphosphate + phosphate
show the reaction diagram
1D-myo-inositol 2,4,5,6-tetrakisphosphate + H2O
1D-myo-inositol 2,4,6-trisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
1D-myo-inositol 2,4,5-trisphosphate + H2O
1D-myo-inositol 2,4-bisphosphate + phosphate
show the reaction diagram
-
preferred substrate
-
-
?
1D-myo-inositol 4,5,6-trisphosphate + H2O
1D-myo-inositol 4,6-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
3-O-methylfluorescein phosphate + H2O
3-O-methylfluorescein + phosphate
show the reaction diagram
-
-
-
-
?
4-nitrophenyl phosphate + H2O
4-nitrophenol + phosphate
show the reaction diagram
-
-
-
-
?
7-methyl-6-thioguanosine + H2O
7-methyl-6-thioguanine + ribose 1-phosphate
show the reaction diagram
-
spectrophotometric continuous coupled enzyme assay substrate
-
-
?
7-nitrobenz-2-oxa-1,3-diazole 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O
7-nitrobenz-2-oxa-1,3-diazole 1-phosphatidyl-1D-myo-inositol 4-phosphate + phosphate
show the reaction diagram
-
fluorescence-labeled substrate
-
-
?
D(+)-sn-1,2-di-O-hexadecanoylglyceryl 1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O
D(+)-sn-1,2-di-O-hexadecanoylglyceryl 1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate + phosphate
show the reaction diagram
D(+)-sn-1,2-di-O-hexadecanoylglyceryl 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O
D(+)-sn-1,2-di-O-hexadecanoylglyceryl 1-phosphatidyl-1D-myo-inositol 4-phosphate + phosphate
show the reaction diagram
D-myo-phosphatidylinositol 3,4,5-trisphosphate
D-myo-phosphatidylinositol 3,4-bisphosphate
show the reaction diagram
D-myo-phosphatidylinositol 3,4,5-trisphosphate
D-myo-phosphatidylinositol 3,4-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
D-myo-phosphatidylinositol 3,5-bisphosphate
D-myo-phosphatidylinositol 3-phosphate
show the reaction diagram
-
-
-
-
?
D-myo-phosphatidylinositol 4,5-bisphosphate
D-myo-phosphatidylinositol 4-phosphate
show the reaction diagram
-
-
-
-
?
D-myo-phosphatidylinositol 4,5-bisphosphate
D-myo-phosphatidylinositol 4-phosphate + phosphate
show the reaction diagram
-
D-myo-phosphatidylinositol 4,5-bisphosphate is a much better substrate than D-myo-phosphatidylinositol 3,4,5-trisphosphate
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-
?
inositol 1,3,4,5-tetrakisphosphate + H2O
inositol 1,3,4-trisphosphate + phosphate
show the reaction diagram
myo-inositol 1,4,5-trisphosphate + H2O
inositol 1,4-diphosphate + phosphate
show the reaction diagram
myo-inositol 3,4,5-trisphosphate + H2O
?
show the reaction diagram
-
-
-
-
?
phosphatidyl-myo-inositol 4,5-bisphosphate + H2O
phosphatidylinositol 4-phosphate + phosphate
show the reaction diagram
phosphatidylinositol 3,4,5-trisphosphate + H2O
phosphatidylinositol 3,4-bisphosphate + phosphate
show the reaction diagram
-
-
?
phosphatidylinositol 3,5-bisphosphate + H2O
phosphatidylinositol 3-phosphate + phosphate
show the reaction diagram
-
-
?
phosphatidylinositol 4,5-bisphosphate + H2O
phosphatidylinositol 4-phosphate + phosphate
show the reaction diagram
-
-
-
-
?
phosphatidylinositol-3,4,5-trisphosphate + H2O
phosphatidylinositol-3,4-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
phosphatidylinositol-4,5-bisphosphate + H2O
phosphatidylinositol-4-phosphate + phosphate
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
1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate + phosphate
show the reaction diagram
1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate + H2O
1-phosphatidyl-1D-myo-inositol 4-phosphate + phosphate
show the reaction diagram
phosphatidylinositol-3,4,5-trisphosphate + H2O
phosphatidylinositol-3,4-bisphosphate + phosphate
show the reaction diagram
-
-
-
-
?
phosphatidylinositol-4,5-bisphosphate + H2O
phosphatidylinositol-4-phosphate + phosphate
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
KCl
-
0.1 M, stimulates
Li+
-
activates
NaCl
-
stimulates
Ni2+
-
activates, competitive binding to other metal ions, KM 0.0064 mM and turnover number 5.48 s-1 at pH 7.0, 30C, recombinant enzyme
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2,3-diphosphoglycerate
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3-benzyl-oxybenzene 1,2,4-trisphosphate
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3-hydroxybenzene 1,2,4-trisphosphate
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4-amino-5-(4-chlorophenyl)-7(t-butyl)pyrazolo(3,4-d)pyrimidine
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inhibits translocation of SHIP1 to the plasma membrane and tyrsine phosphorylation
5,5'-dithiobis(2-nitrobenzoate)
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25-40% inhibition at 0.1 mM, 85-90% inhibition at 5 mM
ammonyx LO
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-
benzene 1,2,3,4-tetrakisphosphate
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-
benzene 1,2,3,5-tetrakisphosphate
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-
benzene 1,2,3-trisphosphate
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-
benzene 1,2,4,5-tetrakisphosphate
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-
benzene 1,2,4-trisphosphate
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-
benzene 1,3,5-trisphosphate
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biphenyl 2,3',4,5',6-pentakisphosphate
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Cetyltrimethylammonium bromide
Cutsum
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-
-
hemoglobin
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at concentrations higher than 1% W/v
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neomycin
-
inhibits only in absence of Triton X-100
o-Iodosobenzoate
-
25-40% inhibition at 0.1 mM, 85-90% inhibition at 5 mM
p-Chloromercuriphenylsulfonate
-
0.1 mM, 98% inhibition
Phenylmercury acetate
-
-
sodium deoxycholate
-
-
Triton X-100
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,2-dioctanoylphosphatidylinositol 4-phosphate
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hydrolysis of D-myo-phosphatidylinositol 4,5-bisphosphate and D-myo-phosphatidylinositol 3,4,5-trisphosphate is stimulated up to 100fold
Cys
-
stimulates
dioctanoylphosphatidylserine
-
-
dithiothreitol
-
stimulates
IRS4
-
binds and activates the enzyme INP51, acts as a negative regulator for 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate contents, IRS4 contains an EPS15 homology domain, interaction analysis
-
mercaptoethanol
-
stimulates
phosphatidylcholine
-
vesicles of phosphatidylcholine only stimulate SHIP2 activity by 2fold. This effect is specific for di-C8 and di-C16 fatty acids of D-myo-phosphatidylinositol 3,4,5-trisphosphate as substrate
phosphatidylserine
-
vesicles of phosphatidylserine (PtdSer) greatly stimulate SHIP2. This effect is specific for di-C8 and di-C16 fatty acids of D-myo-phosphatidylinositol 3,4,5-trisphosphate as substrate
potassium bisperoxo (1,10-phenanthroline) oxovanadate (V)
-
protein phosphatase inhibitor bpV(phen), a cell permeable derivative, enhances the activity of PtdIns3P3 5-phosphatase, particularly SHIP2
reduced glutathione
-
required for full activity
Sodium vanadate
-
protein phosphatase inhibitor enhances the activity of PtdIns3P3 5-phosphatase, particularly SHIP2
TAX4
-
binds and activates the enzyme INP51, acts as a negative regulator for 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphate contents, TAX4 contains an EPS15 homology domain, interaction analysis
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additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.421
1D-myo-inositol 1,4,5-trisphosphate
-
-
0.056
7-methyl-6-thioguanosine
-
pH 7.0, 30C, recombinant enzyme
0.028
Inositol 1,3,4,5-tetrakisphosphate
-
-
0.123
Inositol 1,4,5-trisphosphate
-
-
0.143 - 0.27
phosphatidyl-myo-inositol 4,5-bisphosphate
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
11
7-methyl-6-thioguanosine
-
pH 7.0, 30C, recombinant enzyme
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.068
3-benzyl-oxybenzene 1,2,4-trisphosphate
Homo sapiens
-
-
0.021
3-hydroxybenzene 1,2,4-trisphosphate
Homo sapiens
-
-
0.098
benzene 1,2,3,4-tetrakisphosphate
Homo sapiens
-
-
0.078
benzene 1,2,3,5-tetrakisphosphate
Homo sapiens
-
-
0.086
benzene 1,2,3-trisphosphate
Homo sapiens
-
-
0.004
benzene 1,2,4,5-tetrakisphosphate
Homo sapiens
-
-
0.014
benzene 1,2,4-trisphosphate
Homo sapiens
-
-
0.016
benzene 1,3,5-trisphosphate
Homo sapiens
-
-
0.001
biphenyl 2,3',4,5',6-pentakisphosphate
Homo sapiens
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.044 - 0.11
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-
0.765
-
-
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.7 - 7.3
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-
6.8
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-
6.8 - 8
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7 - 7.4
-
assay at
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.1 - 8.1
-
pH 6.1: about 30% of maximal activity, pH 8.1: about 20% of maximal activity
6.5 - 8.3
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pH 6.5: about 60% of maximal activity, pH 8.3: about 50% of maximal activity
7.2 - 8.4
-
pH 7.2: optimum, pH 8.4: about 65% of maximal activity
additional information
-
pH profile
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37 - 60
-
37C: optimum, 60C: about 27% of maximal activity
additional information
-
activity declines sharply at temperatures above 37C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
high expression of FRA3
Manually annotated by BRENDA team
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by cDNA microarray analysis it is shown that (PI[4,5]P2) phosphatase mRNAas are up-regulated in hepatocytes cultured on a basement membrane matrix, Engelbreth-Holm-Swarm (EHS) gel, which led to the finding that the PI(4,5)P2 levels of hepatocytes decreased on EHS gel
Manually annotated by BRENDA team
-
highly expressed in neurons of the arcuate and lateral nuclei of the hypothalamus. Treatment with a phosphorthioate-modified antisense oligonucleotide for 5ptase IV reduces the hypothalamic expression of 5ptase IV by approximately 80%
Manually annotated by BRENDA team
very low expression of 5PTase14
Manually annotated by BRENDA team
-
murine RAW264.7 macrophages are used
Manually annotated by BRENDA team
-
lowest expression
Manually annotated by BRENDA team
-
highest expression
Manually annotated by BRENDA team
high expression of 5PTase14
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
multicolor total internal reflection fluorescence microscopy (TIRFM) is used to visualize the spatial-temporal recruitment of SJ1, and its binding partner endophilin to clathrin-coated-pits. Differential temporal recruitment of the two major SJ1 splice variants to clathrin-coated-pits is observed. SJ1-145 is rapidly recruited as a burst, together with its binding partner endophilin, at a late stage of clathrin-coated-pit formation; multicolor total internal reflection fluorescence microscopy (TIRFM) is used to visualize the spatialtemporal recruitment of SJ1, and its binding partner endophilin to clathrin-coated-pits. Differential temporal recruitment of the two major SJ1 splice variants to clathrin-coated-pits is observed. SJ1-170 is present at all stages of CCP formation
Manually annotated by BRENDA team
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SKIP localizes to the ER in HuH-7 cells
Manually annotated by BRENDA team
-
dOCRL is associated with endosomes
Manually annotated by BRENDA team
-
cytoplasmic Golgi membrane
Manually annotated by BRENDA team
-
enzyme is associated with the lysosomal membrane
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
in HuH-7 cells overexpressing SKIP and infected with hepatitis B virus (HBV), SKIP localizes to nucleus in addition to endoplasmic reticulum and suppresses HBV gene expression and replication. SKIP loses its nuclear localization and suppressive effect during replication of a core-negative HBV mutant
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36500
-
deduced from cDNA
57000
-
x * 57000, formation of larger aggregates in the absence of salt, SDS-PAGE
72000
-
x * 72000, SDS-PAGE and deduced from gene sequence
100000
-
x * 100000, SDS-PAGE
104000
-
gel filtration
114000 - 120000
-
gel filtration
115000
-
x * 115000, SDS-PAGE
145000
-
-
170000
-
-
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method; sitting drop vapor diffusion method
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
half-life: about 1 day
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
inactivated by freezing
-
Triton X-100 stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 0.1 M NaCl, very littly loss of activity after several months
-
4C, half-life: about 1 day
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
immobilized metal ion affinity chromatography (Co2+), gel filtration
-
Ni2+-colums
-
partial
-
purification by gel filtration chromatography on Sephacryl S-100 followed by cation exchange chromatography on SP-Sepharose
-
recombinant GST-tagged catalytic domains from Escherichia coli strain DH5alpha by glutathione affinity chromatography
recombinant GST-tagged catalytic domains of synaptojanin2 and OCRL from Escherichia coli strain DH5alpha by glutathione affinity chromatography
-
recombinant His-tagged enzyme from yeast by nickel affinity chromatography
recombinant His-tagged full length enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant tagged enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
72-5ptase is expressed as a hemagglutinin (HA) tagged fusion protein in 3T3-L1 cells. Overexpression in adipocytes generates phosphatidylinositol 3-phosphate at the plasma membrane of unstimulated 3T3-L1 adipocytes (conditions under which phosphatidylinositol 3,4,5-triphosphate is not synthesized)
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DNA and amino acid sequence detremination and analysis, expression of GFP-tagged synaptojanin-1 in CHO cells
-
expressed as a HIS-tagged fusion protein in 3T3-L1 adipocytes and in CHO-IR cells (Chinese hamster ovary cells stably transfected with the insulin receptor)
-
expressed in COS-7 cells and in Saccharomyces cerevisiae
-
expressed in Dictyostelium discoideum
-
expressed in Escherichia coli as a GST-fusion protein and as a Flag-tagged fusion protein in human hepatoma HuH-7 cells
-
expressed in Escherichia coli as a GST-tagged protein
-
expressed in HEK293 cells
-
expressed in murine 3T3-L1 preadipocytes
-
expressed in Saccharomyces cerevisiae and in NRK- and HeLa cells
expression in baculovirus-infected Sf9 cells
-
expression in COS7 cells
-
expression of GST-tagged catalytic domain comprising residues A305-A721, in Escherichia coli strain DH5alpha
-
expression of GST-tagged catalytic domain comprising residues S470-R962, in Escherichia coli strain DH5alpha
-
expression of GST-tagged catalytic domains, comprising residues D474-R959 of synaptojanin2 and residues V202-E618 of OCRL, in Escherichia coli strain DH5alpha
-
expression of His-tagged full length enzyme in Escherichia coli strain BL21(DE3)
-
expression of tagged enzyme
-
from oocytes and fertilized eggs, recombinant overexpression of FLAG-tagged proline-rich inositol polyphosphate 5-phosphatase in eggs
-
full-length 5PTase13-encoding sequence is N-terminally fused to a polyhistidinetag and a T7-tag and expressed in Escherichia coli cells
full-length human INPP5B cDNA is cloned into pEGFP-C3 with an N-terminal FLAG tag for in vivo expression. INPP5B cDNA is cloned into pGBKT7 for yeast two-hybrid experiments and pBAC2 for expression of recombinant protein in insect cells. DNA encoding the N-terminal 229 amino acids of INPP5B is cloned into pET41EK-LIC for expression of GST-tagged protein in Escherichia coli
-
full-length SJ1-145 is cloned into the peGFP-C1 vector; full-length SJ1-170 is cloned into the peGFP-C1 vector
-
gene 5PTase14, DNA and amino acid sequence determination and analysis, genetic organization, expression of His-tagged full length enzyme in yeast
gene fra3, expression pattern, DNA and amino acid sequence determination and analysis, complementation of an enzyme-deficient mutant
IPPRp is cloned into the pET17b expression vector and moved into Escherichia coli strain BL21(DE3)pLys S for protein production
-
overexpression of SHIP2 in 3T3-L1 adipocytes, B lymphocytes and L6 myotubes
-
overexpression of SHIP2 in CHO cells
-
overexpression of SHIP2 in COS-7 cells
-
SH2 domain; SH2 domain, a maltose-binding protein fusion protein and a His-tagged version expressed in Escherichia coli
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
enzyme expression, which is markedly elevated during cell differentiation
-
isoform INPP5E is overexpressed in cervical cancer, non-Hodgkins lymphoma, and uterine leiomyosarcoma
-
SHIP2 mRNA and protein expression levels are significantly increased in the brain of type 2 diabetic db/db mice
SKIP expression is markedly elevated during differentiation and is controlled by MyoD in C2C12 cells
-
the enzyme is downregulated in human melanoma
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
DELTA1-465
-
the production of mutant DELTA466-787 containing the catalytic 5-phosphatase domain in DDd5P4 suppresses the growth of wild-type Legionella pneumophila, while the production of DELTA1-465 lacking the catalytic 5-phosphatase domain in Dd5P4-deficient host cells does not affect intracellular growth of the bacteria, indicating that catalytic activity of Dd5P4 is required to attenuate intracellular replication of Legionella pneumophila
DELTARhoGAPDd5P4
-
transfection of dd5p4-null cells with a truncated Dd5P4 lacking the C-terminal RhoGAP domain does not restore endocytic defects, suggesting that the C-terminal RhoGAP domain is essential
C383S
-
mutant with an almost entirely eliminated 3-phosphatase and 4-phosphatase activity, but a maintained 5-phosphatase activity also fails to rescue the endocytic defects in synj1-/- neurons
C392S
-
site-directed mutagenesis, inactivating mutation in the conserved CX5R(T/S) motif of the Sac domain
C641A
-
site-directed mutagenesis, the mutation removes the prenylation site of the enzyme
C910A
-
mutation of the C-terminal cysteine (C910A), which abolishes prenylation of INPP5B does not affect cellular targeting of INPP5B
D192A
-
to assess the role of catalytic activity of SKIP on its suppressive effect, a phosphatase-negative mutant D192A is generated: 5-phosphatase activity is not required for the suppressive effect
D193/E195A
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binding activity is much lower compared to wild-type fragment consisting of amino acids 124-314. Binding activity for active RhoA is dramatically reduced compared to wild-type
D223/K224A
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binding activity is much lower compared to wild-type fragment consisting of amino acids 124-314
D524A
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mutation of the aspartic acid within the conserved sequence PAWCDRIL in the 5-phosphatase domain which renders INPP5B catalytically inactive, has no effect on the targeting of INPP5B to the Golgi apparatus, ERGIC or endosomes
D556A
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site-directed mutagenesis, inactivating point mutation of the conserved DRVL motif of INPP5E
D730A
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mutant with a deficient 5-phosphatase activity fails to rescue the endocytic defects in synj1-/- neurons
DELTA1-885/1186-1258
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the effects of SHIP2 on c-Jun NH2-terminal kinase (JNK) activity and JIP1 (JNK-interacting protein 1) tyrosine phosphorylation are independent of the SHIP2 phosphoinositide 5-phosphatase activity, as similar results are obtained when using a SHIP2 catalytic inactive mutant instead of wild-type SHIP2
DELTA237-893
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the N-terminal domain of OCRL1 is localized throughout the host cytoplasm and it binds Legionella pneumophila LpnE, a Sel1-like repeat protein involved in Legionella-containing vacuoles formation, which localizes to Legionella-containing vacuoles and selectively binds PtdIns(3)P
DELTA73-77
deletion of this LIDIA sequence motif within the N-terminal region of OCRL1 reveals that this sequence motif functions as a second clathrin binding domain in both isoforms
DELTAPIP2
expression of OCRL1 isoform a, but not isoform b, lacking the 5-phosphatase domain impairs transferrin endocytosis
G664D
G664D mutation shows little effect on binding to clathrin, alpha-adaptin, Rac1 or APPL1, while binding to Rabs 5 and 6 is almost completely abolished. The effects of G664D mutation are the same for both OCRL1 isoforms
P105E
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Glu in SHIP1 SH2 domain, faster association kinetics for binding to immobilized peptide VApYSYL
P686A/D690A/R691A
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catalytically inactive dominant-negative mutant inhibits proliferation of preadipocytes more potently than wild-type SHIP2. Phospho-Akt, phospho-ERK1/2, and PDGF receptor (PDGFR) levels are reduced in mutant-expressing preadipocytes. The inhibition of PDGF-activated mitogenic pathways by SHIP2 mutant is consistent with a decrease in PDGFR phosphorylation caused by a drop in receptor levels in SHIP2 mutant-expressing cells. SHIP2 mutant promotes ubiquitination of the PDGFR and its degradation via the lysosomal pathway independently of the association between the E3 ubiquitin ligase c-Cbl and PDGFR
P88S
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Ser in SHIP1 SH2 domain, faster association kinetics for binding to immobilized peptide VApYSYL
D480N
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catalytically inactive 75-5ptase
P687A/D691A/R692G
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liver-specific expression of a dominant-negative SHIP2 mutant in hyperglycemic and hyperinsulinemic KKAy mice increases basal and insulin-stimulated Akt phosphorylation. Protein levels of glucose-6-phosphatase and phosphoenolpyruvate carboxykinase are reduced, and liver produces less glucose through gluconeogenesis. SHIP2 inhibition improves hepatic glycogen metabolism by modulating the phosphorylation states of glycogen phosphorylase and glycogen synthase, which increases hepatic glycogen content. Enhanced glucokinase and reduced pyruvate dehydrogenase kinase 4 expression, together with increased plasma triglycerides, indicate improved glycolysis. Liver-specific inhibition of SHIP2 improves glucose tolerance and markedly reduces prandial blood glucose levels in KKAy mice
E597Q
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site-directed mutagenesis, the mutant enzyme shows an over 140fold increased catalytic efficiency and a 2.4fold reduced affinity for Mg2+ compared to the wild-type enzyme
additional information
APPLICATION
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UNIPROT
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molecular biology
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