BRENDA - Enzyme Database

Localization, purification, and functional reconstitution of the P4-ATPase Atp8a2, a phosphatidylserine flippase in photoreceptor disc membranes

Coleman, J.A.; Kwok, M.C.; Molday, R.S.; J. Biol. Chem. 284, 32670-32679 (2009)

Data extracted from this reference:

Activating Compound
EC Number
Activating Compound
Commentary
Organism
Structure
7.6.2.1
additional information
no activation by phosphatidylcholine or other membrane lipids
Bos taurus
7.6.2.1
phosphatidylethanolamine
activates to a lesser degree
Bos taurus
7.6.2.1
phosphatidylserine
-
Bos taurus
Cloned(Commentary)
EC Number
Commentary
Organism
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression in HEK-293 cells
Homo sapiens
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression in HEK-293 cells
Mus musculus
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression of 1D4-tagged Atp8a2 in HEK-293 cells
Bos taurus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
7.6.2.1
additional information
no inhibition by ouabain and azide
Bos taurus
7.6.2.1
NEM
over 90% inhibition of ATPase activity at 1 mM
Bos taurus
7.6.2.1
vanadate
over 90% inhibition of ATPase activity at 0.1 mM
Bos taurus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
7.6.2.1
additional information
-
additional information
Michaelis-Menten kinetics, overview
Bos taurus
7.6.2.1
additional information
-
additional information
kinetics, overview
Homo sapiens
7.6.2.1
0.704
-
ATP
pH 7.5, 37C
Homo sapiens
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
7.6.2.1
membrane
-
Bos taurus
16020
-
7.6.2.1
membrane
-
Homo sapiens
16020
-
7.6.2.1
membrane
-
Mus musculus
16020
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Bos taurus
-
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Homo sapiens
-
-
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
7.6.2.1
Mg2+
required
Bos taurus
7.6.2.1
Mg2+
required
Homo sapiens
7.6.2.1
Mg2+
required
Mus musculus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
7.6.2.1
132000
-
x * 132000, recombinant enzyme, SDS-PAGE
Bos taurus
7.6.2.1
132000
-
x * 132000, about, recombinant enzyme, SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
Bos taurus
-
ADP + phosphate + phosphatidylethanolamine/out
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
Bos taurus
-
ADP + phosphate + phosphatidylserine/out
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
Mus musculus
-
ADP + phosphate + phospholipid/out
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
Homo sapiens
-
ADP + phosphate + phospholipid/out
-
-
?
7.6.2.1
additional information
Mus musculus
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
7.6.2.1
additional information
Homo sapiens
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
7.6.2.1
additional information
Bos taurus
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
7.6.2.1
Bos taurus
C7EXK4
gene atp8a2
-
7.6.2.1
Homo sapiens
Q9NTI2
gene atp8a2
-
7.6.2.1
Mus musculus
P98200
C57/B6 and BALB/c mice, gene atp8a2
-
Purification (Commentary)
EC Number
Commentary
Organism
7.6.2.1
native Atp8a2 1887fold from photoreceptor outer segments by immunoaffinity chromatography
Bos taurus
7.6.2.1
recombinant enzyme from HEK-293 cells by immunoaffinity chromatography
Homo sapiens
Renatured (Commentary)
EC Number
Commentary
Organism
7.6.2.1
purified Atp8a2 is reconstituted into liposomes containing fluorescent-labeled phosphatidylserine to measure the ability of Atp8a2 to flip phosphatidylserine across the lipid bilayer. 30% of the reconstituted Atp8a2 is inaccessible to trypsin and correspondingly ATP
Bos taurus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Bos taurus
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Homo sapiens
-
7.6.2.1
photoreceptor cell
disc membranes
Bos taurus
-
7.6.2.1
photoreceptor cell
disc membranes
Homo sapiens
-
7.6.2.1
photoreceptor cell
disc membranes
Mus musculus
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells, high expression of atp8a2
Bos taurus
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells, high expression of atp8a2
Homo sapiens
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells
Mus musculus
-
7.6.2.1
testis
high expression of atp8a2
Bos taurus
-
7.6.2.1
testis
high expression of atp8a2
Homo sapiens
-
7.6.2.1
testis
-
Mus musculus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
7.6.2.1
0.054
-
purified native enzyme, pH 7.5, 37C
Bos taurus
7.6.2.1
35
-
purified native enzyme reconstituted in lipid vesicles, pH 7.5, 37C
Bos taurus
7.6.2.1
45
55
purified lipid-stimulated enzyme, pH 7.5, 37C
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
-
712395
Bos taurus
ADP + phosphate + phosphatidylethanolamine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
the enzyme is specific for ATP and shows no activity with GTP, AMP-PNP, GDP, or ADP
712395
Bos taurus
ADP + phosphate + phosphatidylethanolamine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
-
712395
Bos taurus
ADP + phosphate + phosphatidylserine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
the enzyme is specific for ATP and shows no activity with GTP, AMP-PNP, GDP, or ADP
712395
Bos taurus
ADP + phosphate + phosphatidylserine/out
-
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
-
712395
Mus musculus
ADP + phosphate + phospholipid/out
-
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
-
712395
Homo sapiens
ADP + phosphate + phospholipid/out
-
-
-
?
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Mus musculus
?
-
-
-
-
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Homo sapiens
?
-
-
-
-
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Bos taurus
?
-
-
-
-
7.6.2.1
additional information
dithionite NBD-lipid assay for detection of flippase activity
712395
Homo sapiens
?
-
-
-
-
7.6.2.1
additional information
dithionite NBD-lipid assay for detection of flippase activity
712395
Bos taurus
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
7.6.2.1
?
x * 132000, recombinant enzyme, SDS-PAGE
Bos taurus
7.6.2.1
?
x * 132000, about, recombinant enzyme, SDS-PAGE
Homo sapiens
Temperature Optimum [C]
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
7.6.2.1
23
-
flippase assay at
Bos taurus
7.6.2.1
23
-
flippase assay at
Homo sapiens
7.6.2.1
37
-
ATPase assay at
Bos taurus
7.6.2.1
37
-
ATPase assay at
Homo sapiens
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6.2.1
7
9
with substrate phosphatidylserine
Bos taurus
7.6.2.1
7.5
-
assay at
Homo sapiens
7.6.2.1
8
-
with substrate phosphatidylethanolamine
Bos taurus
Activating Compound (protein specific)
EC Number
Activating Compound
Commentary
Organism
Structure
7.6.2.1
additional information
no activation by phosphatidylcholine or other membrane lipids
Bos taurus
7.6.2.1
phosphatidylethanolamine
activates to a lesser degree
Bos taurus
7.6.2.1
phosphatidylserine
-
Bos taurus
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression in HEK-293 cells
Homo sapiens
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression in HEK-293 cells
Mus musculus
7.6.2.1
gene atp8a2, DNA and amino acid sequence determination and analysis, expression of 1D4-tagged Atp8a2 in HEK-293 cells
Bos taurus
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
7.6.2.1
additional information
no inhibition by ouabain and azide
Bos taurus
7.6.2.1
NEM
over 90% inhibition of ATPase activity at 1 mM
Bos taurus
7.6.2.1
vanadate
over 90% inhibition of ATPase activity at 0.1 mM
Bos taurus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
7.6.2.1
additional information
-
additional information
Michaelis-Menten kinetics, overview
Bos taurus
7.6.2.1
additional information
-
additional information
kinetics, overview
Homo sapiens
7.6.2.1
0.704
-
ATP
pH 7.5, 37C
Homo sapiens
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
7.6.2.1
membrane
-
Bos taurus
16020
-
7.6.2.1
membrane
-
Homo sapiens
16020
-
7.6.2.1
membrane
-
Mus musculus
16020
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Bos taurus
-
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Homo sapiens
-
-
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
7.6.2.1
Mg2+
required
Bos taurus
7.6.2.1
Mg2+
required
Homo sapiens
7.6.2.1
Mg2+
required
Mus musculus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
7.6.2.1
132000
-
x * 132000, recombinant enzyme, SDS-PAGE
Bos taurus
7.6.2.1
132000
-
x * 132000, about, recombinant enzyme, SDS-PAGE
Homo sapiens
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
Bos taurus
-
ADP + phosphate + phosphatidylethanolamine/out
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
Bos taurus
-
ADP + phosphate + phosphatidylserine/out
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
Mus musculus
-
ADP + phosphate + phospholipid/out
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
Homo sapiens
-
ADP + phosphate + phospholipid/out
-
-
?
7.6.2.1
additional information
Mus musculus
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
7.6.2.1
additional information
Homo sapiens
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
7.6.2.1
additional information
Bos taurus
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
7.6.2.1
native Atp8a2 1887fold from photoreceptor outer segments by immunoaffinity chromatography
Bos taurus
7.6.2.1
recombinant enzyme from HEK-293 cells by immunoaffinity chromatography
Homo sapiens
Renatured (Commentary) (protein specific)
EC Number
Commentary
Organism
7.6.2.1
purified Atp8a2 is reconstituted into liposomes containing fluorescent-labeled phosphatidylserine to measure the ability of Atp8a2 to flip phosphatidylserine across the lipid bilayer. 30% of the reconstituted Atp8a2 is inaccessible to trypsin and correspondingly ATP
Bos taurus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Bos taurus
-
7.6.2.1
additional information
localization study by immunofluorescence microscopy
Homo sapiens
-
7.6.2.1
photoreceptor cell
disc membranes
Bos taurus
-
7.6.2.1
photoreceptor cell
disc membranes
Homo sapiens
-
7.6.2.1
photoreceptor cell
disc membranes
Mus musculus
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells, high expression of atp8a2
Bos taurus
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells, high expression of atp8a2
Homo sapiens
-
7.6.2.1
retina
outer segment disc membranes of rod and cone photoreceptor cells
Mus musculus
-
7.6.2.1
testis
high expression of atp8a2
Bos taurus
-
7.6.2.1
testis
high expression of atp8a2
Homo sapiens
-
7.6.2.1
testis
-
Mus musculus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [mol/min/mg]
Specific Activity Maximum [mol/min/mg]
Commentary
Organism
7.6.2.1
0.054
-
purified native enzyme, pH 7.5, 37C
Bos taurus
7.6.2.1
35
-
purified native enzyme reconstituted in lipid vesicles, pH 7.5, 37C
Bos taurus
7.6.2.1
45
55
purified lipid-stimulated enzyme, pH 7.5, 37C
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
-
712395
Bos taurus
ADP + phosphate + phosphatidylethanolamine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylethanolamine/in
the enzyme is specific for ATP and shows no activity with GTP, AMP-PNP, GDP, or ADP
712395
Bos taurus
ADP + phosphate + phosphatidylethanolamine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
-
712395
Bos taurus
ADP + phosphate + phosphatidylserine/out
-
-
-
?
7.6.2.1
ATP + H2O + phosphatidylserine/in
the enzyme is specific for ATP and shows no activity with GTP, AMP-PNP, GDP, or ADP
712395
Bos taurus
ADP + phosphate + phosphatidylserine/out
-
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
-
712395
Mus musculus
ADP + phosphate + phospholipid/out
-
-
-
?
7.6.2.1
ATP + H2O + phospholipid/in
-
712395
Homo sapiens
ADP + phosphate + phospholipid/out
-
-
-
?
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Mus musculus
?
-
-
-
-
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Homo sapiens
?
-
-
-
-
7.6.2.1
additional information
Atp8a2 flipps fluorescent-labeled phosphatidylserine from the inner leaflet of liposomes, equivalent to the exocytoplasmic leaflet of cell membranes, to the outer leaflet, equivalent to cytoplasmic leaflet, in an ATP-dependent manner
712395
Bos taurus
?
-
-
-
-
7.6.2.1
additional information
dithionite NBD-lipid assay for detection of flippase activity
712395
Homo sapiens
?
-
-
-
-
7.6.2.1
additional information
dithionite NBD-lipid assay for detection of flippase activity
712395
Bos taurus
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
7.6.2.1
?
x * 132000, recombinant enzyme, SDS-PAGE
Bos taurus
7.6.2.1
?
x * 132000, about, recombinant enzyme, SDS-PAGE
Homo sapiens
Temperature Optimum [C] (protein specific)
EC Number
Temperature Optimum [C]
Temperature Optimum Maximum [C]
Commentary
Organism
7.6.2.1
23
-
flippase assay at
Bos taurus
7.6.2.1
23
-
flippase assay at
Homo sapiens
7.6.2.1
37
-
ATPase assay at
Bos taurus
7.6.2.1
37
-
ATPase assay at
Homo sapiens
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
7.6.2.1
7
9
with substrate phosphatidylserine
Bos taurus
7.6.2.1
7.5
-
assay at
Homo sapiens
7.6.2.1
8
-
with substrate phosphatidylethanolamine
Bos taurus
General Information
EC Number
General Information
Commentary
Organism
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Bos taurus
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Homo sapiens
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Mus musculus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Bos taurus
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Homo sapiens
7.6.2.1
physiological function
P4-ATPases in general are implicated in the energy-dependent translocation of aminophospholipids across cell membranes. Atp8a2 is implicated in the generation and maintenance of phosphatidylserine asymmetry in photoreceptor disc membranes
Mus musculus