Information on EC 3.6.1.3 - adenosinetriphosphatase

Word Map on EC 3.6.1.3
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.6.1.3
-
RECOMMENDED NAME
GeneOntology No.
adenosinetriphosphatase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O = ADP + phosphate
show the reaction diagram
(Ca2+-Mg2+)-ATPase: no phosphorylated intermediate formed
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
NIL
-
-
Purine metabolism
-
-
purine metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase
Many enzymes previously listed under this number are now listed separately under EC 3.6.3 and EC 3.6.4.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
strain KM
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain MSR-1
UniProt
Manually annotated by BRENDA team
young Swiss strain male albino mice
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
strain OT3
-
-
Manually annotated by BRENDA team
strain OT3
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
SwissProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
additional information
-
zoledronate treatment promotes F1-expression as well as endogenous phosphoantigen production. Recognition of phosphoantigens on cell membranes in the form of nucleotide derivatives that can bind to F1-ATPase acting as a presentation molecule
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + H2O
ADP + phosphate
show the reaction diagram
ATP + H2O
AMP + diphosphate
show the reaction diagram
-
-
-
-
?
CTP + H2O
CDP + phosphate
show the reaction diagram
dATP + H2O
dADP + phosphate
show the reaction diagram
dCTP + H2O
dCDP + phosphate
show the reaction diagram
-
-
-
-
?
GTP + H2O
GDP + phosphate
show the reaction diagram
ITP + H2O
IDP + phosphate
show the reaction diagram
UTP + H2O
UDP + 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
ATP + H2O
ADP + phosphate
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
ATP
-
protease La is an ATP-dependent protease, it requires ATP hydrolysis to digest larger, intact proteins, but can cleave small, fluorogenic peptides such as Glu-Ala-Ala-Phe-MNA by only binding, but not hydrolyzing, ATP
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
(NH4)2SO4
-
the enzyme requires NaCl for maximal activity but Na2SO4 and (NH4)2SO4 can substitute. Five times more activity than in the presence of NaCl is observed when 1 M (NH4)2SO4 is present
bafilomycin A1
-
0.001 mM, 64% inhibition
Co2+
Co2+ gives 70% of the activity measured with Mg2+
Dicyclohexylcarbodiimide
-
0.3 mM, 59% inhibition
KNO3
-
100 mM, 14% inhibition
N-ethylmaleimide
-
1 mM, 5% inhibition
Na2SO4
-
the enzyme requires NaCl for maximal activity but Na2SO4 and (NH4)2SO4 can substitute. Sodium sulfate (1.5 M) can substitute to an extent for NaCl resulting in 80% of original enzyme activity
Na3VO4
-
0.1 mM, 32% inhibition
NaCl
-
maximal enzyme activity in the presence of 2.5 M NaCl. The enzyme requires NaCl for maximal activity but Na2SO4 and (NH4)2SO4 can substitute
NaN3
-
5 mM, complete inhibition. Azide can bind via the Mg2+-binding sites (whether ATP is in the active site or not) and inactivate the enzyme. It is a mixed non-competitive inhibitor because it binds more readily to the enzyme than the enzyme/substrate complex
oubain
-
15 mM, complete inhibition
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4,5-dihydro-8H-6-(N-decyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 30-100 mg/l
4,5-dihydro-8H-6-(N-dodecyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 1-3 mg/l
4,5-dihydro-8H-6-(N-dodecylamino)-1-(2'-deoxy-alpha-D-erythropentofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 3-10 mg/l
4,5-dihydro-8H-6-(N-dodecylamino)-1-(2'-deoxy-beta-D-erythropentofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 3-10 mg/l
4,5-dihydro-8H-6-(N-hexadecyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 250 mg/l
4,5-dihydro-8H-6-(N-octadecyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 5.0 mg/l
4,5-dihydro-8H-6-(N-tetradecyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
IC50: 3-10 mg/l
4-chloro-7-nitrobenzofurazan
-
-
5-fluoro-2-selenocytosine
-
IC50: 0.075 mM for NTPase reaction, no influence to helicase activity up to a concentration of 0.5 mM
7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole
-
-
aflatoxin
-
obtained from growing Aspergillus parasiticus in SMKY liquid medium, inhibits enzyme activity at high doses in vivo in testis, liver and kidney, curcumin along with aflatoxin ameliorates aflatoxin-induced changes in adenosine triphosphatase activities
ATP
-
substrate inhibition of wild-type and mutant enzymes, kinetics, overview
Ca2+
-
solubilized enzyme, 359% of initial acitivity in presence of 1 mM. For pH value above 6.5, inhibitory above 2.5 mM
CaCl2
in presence of Mn2+, 10 mM MgCl2 inhibits 20%
Co2+
-
2.5 mM, 61% residual activity
EDTA
-
0.1 mM, complete loss of activity
etretinate
-
-
HCl
-
0.1 M, irreversible inactivation
K+
-
above 500 mM
KNO3
-
20 mM, 63% inhibition
MgCl2
in presence of Mn2+, 10 mM MgCl2 inhibits 10%
Mn2+
-
2.5 mM, 78% residual activity
N,N'-dicyclohexylcarbodiimide
N-Ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
-
-
N-ethylmaleimide
Na+
-
above 500 mM
NaCl
100 mM, inhibits ATP hydrolysis by 30%
NaN3
-
the enzyme is non-competitively inhibited
Ni2+
-
2.5 mM, 18% residual activity
nitrate
NO3-
-
50% inhibition at 8 mM, maximal inhibition of 87% at 50 mM
O6-benzyl-N7-chloroethylguanine
-
weak inhibition of NTPase activity, enhances helicase activity
O6-benzylguanine
-
weak inhibitor of ATPase and helicase activity
oligo(dA)25
-
above 0.5 mM, inhibits unwinding reaction
-
p-chloromercuribenzenesulfonate
-
-
p-Chloromercuriphenyl sulfonate
-
500 nM at 35% inhibition. Inhibition is reversed by cysteine
p-hydroxymercuribenzoate
-
-
polyA
inhibits to a lesser extent than polyC and polyU
polyC
1 mg/l, 70% inhibition
polyG
inhibits to a lesser extent than polyC and polyU
polyU
1 mg/l, 70% inhibition
quercetin
retinoic acid
-
-
ribavirin-TP
-
at ATP concentration equal to Km, IC50 of NTPase reaction is 0.4 mM, classical competitive inhibitor with regard to ATP. At ATP and DNA duplex concentrations corresponding to their KM-values an IC50 of 0.12 mM is measured. Inhibition reaches a maximum of 30% of the control at 0.45 mM and is not competitive with regard to ATP
RNA
homopolymeric RNA inhibits under otherwise optimal conditions
SDS
-
inactivation of the enzyme at 12% SDS
SO42-
-
50% inhibition at 25 mM
Sodium azide
-
-
sodium orthovanadate
-
-
sulfate
-
in the presence of 50 mM sulfate, the ATPase activity at pH 5.5 is inhibited by about 33%, whereas at pH 7 the activity is reduced only about 15%
vanadate
-
no inhibitory effect on the ATPase activity at pH 7.0, whereas a remarkable inhibition at high concentrations can be observed for the activity at pH 5.5
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4,5-dihydro-8H-6-(N-phenyl)amino-1-(beta-D-ribofuranosyl)imidazo[4,5-e][1,3]diazepine-4,8-dione
-
activates
Calmodulin
-
-
disodium carbonate
-
20 mM, 6.8fold stimulation at pH 7.0
dithiothreitol
-
2 mM, 1.9fold activation
HCO3-
-
slight inhibition
Na2SO3
-
20 mM, 10.5fold stimulation at pH 7.0, 3.3fold stimulation at pH 5.0; 20 mM, 3fold stimulation at pH 5.0
O6-benzyl-N7-chloroethylguanine
-
weak inhibition of NTPase activity, enhances helicase activity up to 850% of control
O6-benzyl-N9-chloroethylguanine
-
stimulates NTPase activity with a maximum effect of 350% of control at 0.65 mM, enhances helicase activity up to 220% of control
poly(dA)
-
1.7-3.3 mM, activation of ATPase activity to 170-180% of control
ssDNA
sulfate
-
10 mM, activating with shift of pH optimum from 6.5 to 5.0
sulfite
triphosphoric acid 1-adenosin-5'-yl ester 3-(3-methylbut-3-enyl) ester
-
an adenylated derivative of isopentenyl diphosphate, can stably bind to F1-ATPase-coated beads and promotes TCR aggregation, lymphokine secretion, and activation of the cytolytic process provided that nucleotide diphosphatase activity is present. It also acts as an allosteric activator of F1-ATPase
verapamil
-
ATPase activity is stimulated by verapamil
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 4
ATP
0.0066
dATP
-
37C
0.071
dCTP
-
37C
0.0000000047
DNA duplex
-
-
-
0.002
GTP
-
37C
additional information
additional information
-
pre-steady-state and steady-state kinetics, functional nonequivalency in the ATPase activity of the enzyme that contains high- and low-affinity ATPase sites, which are noninteracting, overview, ATP and peptide hydrolysis are not stoichiometrically linked
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0009 - 133
ATP
1.56
dATP
Reovirus sp.
-
37C
0.022
dCTP
Reovirus sp.
-
37C
0.088
GTP
Reovirus sp.
-
37C
additional information
additional information
West Nile virus
-
at optimum Mg2+ and saturating ATP concentrations 1 pmol of enzyme unwinds 5.5 fmol of DNA duplex per s
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.001 - 1.4
ATP
4
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.1
NaN3
-
pH 7.8, 37C, in the presence of Na+, K+, and Mg2+
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.075
5-fluoro-2-selenocytosine
West Nile virus
-
IC50: 0.075 mM for NTPase reaction, no influence to helicase activity up to a concentration of 0.5 mM
0.025
erythrosin B
Sulfolobus acidocaldarius
-
pH 6.0, 70C
0.4
N-Ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline
Sulfolobus acidocaldarius
-
pH 6.0, 70C
8
nitrate
Sulfolobus acidocaldarius
-
pH 6.0, 70C
0.17
p-hydroxymercuribenzoate
Sulfolobus acidocaldarius
-
pH 6.0, 70C
0.12
ribavirin-TP
West Nile virus
-
at ATP concentration equal to Km, IC50 of NTPase reaction is 0.4 mM, classical competitive inhibitor with regard to ATP. At ATP and DNA duplex concentrations corresponding to their KM-values an IC50 of 0.12 mM is measured. Inhibition reaches a maximum of
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00000034
-
-
0.001
-
below, purified recombinant mutant K939M/K1952M enzyme
0.0583
wild-type strain MSR-1
0.0787
mutant strain NPHB
0.2
-
80C, pH not specified in the publication
0.486
-
pH 6.0, 70C
0.5
-
above, purified recombinant wild-type enzyme
0.566
-
pH 7.5, 70C
5.7
-
pH 5.2, 60C
8.4
-
pH 7.8, 37C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.3
-
second optimum at pH 8.0
5.5 - 7
-
when the ATP:Mg2+ ratio is 4:1. At higher Mg2+-concentrations only one pH-maximum at pH 5.5 can be observed. In the presence of 20 mM Na2SO3 the ATPase activity can be stimulated 2.5fold in the pH range from 5.5 to 7.0
5.5
-
second optimum at pH 8.0
6.25
-
sole optimum in presence of sulfite
7 - 7.5
-
assay at
7.2
-
pH optimum for ratio Ca2+/ATP of 1, or Mg2+/ATP of 1.25
7.2 - 7.5
ATPase activity
additional information
-
optimum pH for the enzyme activity varies depending on the ratio of ATP to Mg2+ or Ca2+
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4 - 6.7
-
55C, pH 4.0: about 70% of maximal activity, pH 6.7: about 40% of maximal activity
6 - 7.6
-
pH 6.0: about 80% of maximal activity, pH 7.6: about 60% of maximal activity
6.8 - 8
pH 6.8: about 90% of maximal activity, pH 8.0: about 70% of maximal activity
7 - 8
pH 7.0: about 80% of maximal activity, pH 8.0: about 55% of maximal activity, mutant enzyme lacking the N-terminal transmembrane helix (residues 127), MBA1DELTAN
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 50
little increase as the temperature is raised from 30C to 50C
37
-
assay at
42 - 50
-
-
75
mutant enzyme lacking the N-terminal transmembrane helix (residues 127), MBA1DELTAN, activity is about a third of wild-type rates
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30 - 70
30C-50C: little increase as the temperature is raised from 30C to 50C, 70C: about 50% of maximal activity
40 - 70
40C: about 55% of maximal activity, 70C: about 80% of maximal activity
50 - 80
-
50C: about 50% of maximal activity, 80C: about 80% of maximal activity
50 - 100
50C: about 50% of maximal activity, 100C: about 70% of maximal activity
65 - 80
65C: about 40% of maximal activity, 80C: about 75% of maximal activity, no activity at room temperature or 37C, no activity beyond 85C. Mutant enzyme lacking the N-terminal transmembrane helix (residues 127), MBA1DELTAN
70 - 95
-
pH 8.0, 70C: about 55% of maximal activity, 95C: about 50% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
from virus-infected Vero cells
Manually annotated by BRENDA team
-
activated by phosphoantigens provided exogenously or produced by tumors and infected cells. Activation requires a contact between Vgamma9Vdelta2 cells and neighboring cells, F1-ATPase is required for the response to inosine diphosphate
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
specific isozyme of V-ATPase
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Aquifex aeolicus (strain VF5)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Deinococcus radiodurans (strain ATCC 13939 / DSM 20539 / JCM 16871 / LMG 4051 / NBRC 15346 / NCIMB 9279 / R1 / VKM B-1422)
Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
Sulfolobus acidocaldarius (strain ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770)
Sulfolobus acidocaldarius (strain ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770)
Sulfolobus acidocaldarius (strain ATCC 33909 / DSM 639 / JCM 8929 / NBRC 15157 / NCIMB 11770)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
24000
-
x * 63000 + x * 48000 + x * 24000, SDS-PAGE
28000
-
3 * 69000 + 3 * 54000 + 1 * 28000, most probable subunit stoichiometry, SDS-PAGE
28090
-
6 * 28090, SDS-PAGE
30414
1 * 30414, mass spectrometry, the enzyme is monomeric in solution in the absence of DNA but forms large molecular mass complexes with predominant peaks at approximately 250000 and 600000 Da when incubated with a 34mer oligonucleotide
30416
1 * 30416, calculated from sequence, the enzyme is monomeric in solution in the absence of DNA but forms large molecular mass complexes with predominant peaks at approximately 250000 and 600000 Da when incubated with a 34mer oligonucleotide
40000
-
6 * 40000, ultracentrifugation measurements, the protein forms ring-shaped complexes with a diameter of 125 A. It assembles into hexamers over a wide concentration range both in presence and absence of ATP
48000
-
x * 63000 + x * 48000 + x * 24000, SDS-PAGE
51000
-
3 * 65000, alpha subunit, + 3 * 51000, beta subunit, SDS-PAGE
54000
-
3 * 69000 + 3 * 54000 + 1 * 28000, most probable subunit stoichiometry, SDS-PAGE
60000
-
x * 60000, SDS-PAGE
61000
-
x * 74000 + x * 61000, SDS-PAGE
63000
-
x * 63000 + x * 48000 + x * 24000, SDS-PAGE
65000
-
3 * 65000, alpha subunit, + 3 * 51000, beta subunit, SDS-PAGE
67000
-
x * 67000, SDS-PAGE, Lon exists as a homo-oligomer and represents one of the simplest ATP-dependent proteases because both the protease and ATPase domains are located within each monomeric subunit
69200
2 * 69200, wild-type enzyme forms a mixture of dimers and higher oligomers in solution. The mutant lacking the N-terminal transmembrane helix (residues 127), MBA1DELTAN forms dimers; 6 * or 7 * 69200, wild-type enzyme forms a mixture of dimers and higher oligomers in solution. The mutant lacking the N-terminal transmembrane helix (residues 127), MBA1DELTAN forms dimers
74000
-
x * 74000 + x * 61000, SDS-PAGE
130000
-
gel filtration
153900
-
recombinant enzyme, dynamic light-scattering
360000
-
gel filtration
370000
-
gel filtration
430000
-
gel filtration
440000 - 480000
gel filtration
additional information
-
molecular weight above 100000 Da, gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heptamer
-
3 * 69000 + 3 * 54000 + 1 * 28000, most probable subunit stoichiometry, SDS-PAGE
hexamer
monomer
oligomer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
vapor diffusion method, using 0.2 M magnesium chloride, 0.1 M Bis-Tris, 25% (w/v) PEG-3350, ADP and MgCl2 at pH 5.5 and 4C (isoform HSPA1A), or 0.2 M trimethyl amine n-oxide, 0.1 M Tris, 26% (w/v) PEG monomethyl ether-2000, ADP and MnCl2, at pH 8.5 and 4C (isoform HSPA1L), or 0.2 M ammonium acetate, 0.1M Bis-Tris, 25% (w/v) PEG-3350, ADP and MgCl2, at pH 5.5 and 4C (isoform HSPA2), or 0.2 M calcium chloride, 0.1 M sodium acetate, 20% (w/v) PEG-6000, ADP and MgCl2, at pH 5.0 and 20C (isoform HSPA5), or 0.1M disodium hydrogen phosphate, 0.1 M citric acid, 16% (w/v) PEG-300, ADP and MgCl2, at pH 3.2 and 20C (isoform HSPA6)
-
hanging-drop vapour-diffusion method, crystals belong to the orthorhombic space group C2, with unit-cell parameters a = 79.41, b = 48.63, c = 108.77 A, and diffract to beyond 2.6 A resolution; sitting-drop vapour-diffusion method, hanging-drop vapour-diffusion method, Crystals belong to the orthorhombic space group C2, with unit-cell parameters a = 79.41, b = 48.63, c = 108.77 A, and diffract to beyond 2.6 A resolution
ATPase PH0284 in complex with ATP, oil-microbatch technique, 0.001 ml of protein solution containing 18.9 mg/ml protein, 200 mM NaCl, and 20 mM Tris-HCl, pH 8.0, is mixed with 0.001 ml of precipitant solution containing 1.5 M ammonium sulfate, 25% v/v glycerol, and 75 mM Tris-HCl, pH 8.5, sealing with a 1:1 mixture of paraffin oil and silicone, 5-6 days at 22C, X-ray diffraction structure determination and analysis at 2.0-2.3 A resolution
-
sitting-drop vapour diffusion at 20C, hanging-drop vapour-diffusion method, high-resolution crystal structure
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
-70
-
cold labile
-20 - 25
-
24 h, the enzyme loses over 50% of its activity
89
-
20 min, no loss of activity
95
-
Arrhenius plots for both membrane bound ATPase activities are linear up to 95C, reflecting the enormous thermostability of the enzyme
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
labile to storage in nitrogen
-
the addition of 30% sorbitol or 20% glycerol is not effective in maintaining activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-19C, 2 days, enzyme loses about 80% of its initial activity
-
4C, stable for at least 3 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
enrichment
-
expressed and purified without any affinity tag by a combination of heat treatment, heparin and gel-filtration column chromatography; XPB2 is expressed with a cleavable N-terminal polyhistidine tag. The XPB2 protein is unstable at temperatures above 50 C, so the tagged protein is purified without a heat step by immobilised metal affinity chromatography followed by chromatography on a heparin column
HiTrap chelating column chromatography and Superdex-200 gel filtration
-
partial, solubilization by dialysis or mild alkali treatment
-
protein can be easily removed from the membrane by mild treatment with zwitterionic detergents
-
recombinant enzyme from Escherichia coli strain BL21(DE3) by anion exchange chromatography, adsorption chromatography, ultrafiltration, and gel filtration to homogeneity
-
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta 2 (DE3) by nickel affinity chromatography and gel filtration
-
recombinant wild-type and mutant His-tagged ABCA1s from insect Sf9 cell microsomal membranes by nickel affinity and anion exchange chromatography
-
release from the membrane by washing with 0.003 M Tris-HCl, pH 7.5
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; expression in Escherichia coli
cloning of gene CHD1L, previously called ALC1, within the 1q21 amplicon, by hybrid selection using microdissected DNA from chromosome 1q21
-
expressed in Escherichia coli
-
expressed in Escherichia coli BL21(DE3)R3 pRARE cells
-
expression analysis of the ATPase gene of wild-type and mutant enzymes
expression in Escherichia coli
expression in Escherichia coli strain BL21(DE3)
-
expression in Escherichia coli; expression in Escherichia coli
expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli; expression in Escherichia coli
expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta 2 (DE3)
-
heterologous expression in Escherichia coli with a C-terminal His6-tag; heterologuous expression in Escherichia coli with a C-terminal His6-tag. The full-length form is recovered from the membrane fraction and the N-terminally deleted form lacking the transmembrane sequence, MBA2DELTAN, from the cytosolic fraction, but part of the latter protein appears to be membrane-associated
overexpressed in Escherichia coli
-
stable expression of ABCA1 in human fibroblasts WI-38 and HEK-293 cell membranes, expression of His-tagged wild-type and mutant ABCA1s in Spodoptera frugiperda Sf9 cells using the baculovirus transfection system
-
the selenomethionine derivative of the full length SSO1545 is expressed in Escherichia coli with an N-terminal TEV cleavable His-tag
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
the protein is induced at 200 J/m2, and the transcription and translation levels are approximately twofold compared to those in the mock. The transcription and translation levels do not increase at the dose of 100 J/m2
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
G893A
-
site-directed mutagenesis, the mutant shows 63% of wild-type ATPase activity, 80% of wild-type protease activity, and 2.27fold of the activation by beta-casein compared to the wild-type enzyme
G893A/G894A
-
site-directed mutagenesis, the mutant shows 103% of wild-type ATPase activity, 79% of wild-type protease activity, and 0.745fold of the activation by beta-casein compared to the wild-type enzyme
G893A/G894P
-
site-directed mutagenesis, the mutant shows 112% of wild-type ATPase activity, no protease activity, and 0.32fold of the activation by beta-casein compared to the wild-type enzyme
G893P
-
site-directed mutagenesis, the mutant shows 89% of wild-type ATPase activity, no protease activity, and 0.49fold of the activation by beta-casein compared to the wild-type enzyme
G893P/G894A
-
site-directed mutagenesis, the mutant shows 71% of wild-type ATPase activity, 8% of wild-type protease activity, and 0.23fold of the activation by beta-casein compared to the wild-type enzyme
G894A
-
site-directed mutagenesis, the mutant shows 139% of wild-type ATPase activity, 76% of wild-type protease activity, and 0.49fold of the activation by beta-casein compared to the wild-type enzyme
G894P
-
site-directed mutagenesis, the mutant shows 130% of wild-type ATPase activity, 84% of wild-type protease activity, and 0.23fold of the activation by beta-casein compared to the wild-type enzyme
G894S
-
site-directed mutagenesis, the mutant shows 140% of wild-type ATPase activity, 47% of wild-type protease activity, and 0.88fold of the activation by beta-casein compared to the wild-type enzyme
K529R
-
site-directed mutagenesis, inactive mutant
K939M/K1952M
-
site-directed mutagenesis of Walker A motif residues, the mutant shows highly reduced activity compared to the wild-type enzyme
S855A
-
site-directed mutagenesis, the mutant shows 78% of wild-type ATPase activity, no protease activity, and 0.35fold of the activation by beta-casein compared to the wild-type enzyme
T880V
-
site-directed mutagenesis, the mutant shows 46% of wild-type ATPase activity, 107% of wild-type protease activity, and 3.28fold of the activation by beta-casein compared to the wild-type enzyme
W770A
-
site-directed mutagenesis, the mutant shows 98.5% of wild-type ATPase activity, 6.4% of wild-type protease activity, and 0.305fold of the activation by beta-casein compared to the wild-type enzyme
W770P
-
site-directed mutagenesis, the mutant shows 123% of wild-type ATPase activity, 55.3% of wild-type protease activity, and 0.64fold of the activation by beta-casein compared to the wild-type enzyme
S154A
-
mutant with negligible ATPase activity
K192A
Walker A box mutant form shows no ATPase activity
K96A
Walker A box mutant form of xpb2 shows no ATPase activity
up
is expressed when the cells are grown on rich medium containing tryptone, sucrose and yeast extract; the expression levels of SSO0120 varies depending on the growth phase. It is highly expressed in the stationary phase of arabinose- and maltose-grown cells and exponentially growing tryptone cells
K192A
-
Walker A box mutant form shows no ATPase activity
-
K96A
-
Walker A box mutant form of xpb2 shows no ATPase activity
-
up
-
is expressed when the cells are grown on rich medium containing tryptone, sucrose and yeast extract; the expression levels of SSO0120 varies depending on the growth phase. It is highly expressed in the stationary phase of arabinose- and maltose-grown cells and exponentially growing tryptone cells
-
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATPase activity in the crude alkaline extract of the membrane can recombine with the alkali-treated membrane in the presence of Ca2+ or Mg2+. Upon recombination, 10-40% of the activity becomes protected from cold inactivation at 0C. The purified ATPases can also recombine with the membrane in the presence of Ca2+, with Mg2+ being much less effective
-
reconstitution of purified recombinant enzyme in lipid vesicles consisting of L-alpha-lecitin from soybean, sphingomyelin, synthesized phospholipids, or sterols, in 40 mM Tris-HCl, pH 7.5, 0.1 mM EGTA, overview
-
solubilized ATPase binds back to membranes depleted of ATPase, in the presence of 0.01 M Ca2+. It does not bind to undepleted membranes
-
Show AA Sequence (5570 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.