Information on EC 3.6.3.16 - arsenite-transporting ATPase

Word Map on EC 3.6.3.16
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 expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
3.6.3.16
-
RECOMMENDED NAME
GeneOntology No.
arsenite-transporting ATPase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + H2O + arsenite/in = ADP + phosphate + arsenite/out
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric ester
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
arsenate detoxification II (glutaredoxin)
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP phosphohydrolase (arsenite-exporting)
A multisubunit non-phosphorylated ATPase that is involved in the transport of ions. A bacterial enzyme that usually contains two subunits where one (with 12 membrane-spanning segments) forms the 'channel' part and the other (occurring in pairs peripherally to the membrane) contains the ATP-binding site. Exports arsenite and antimonite anions.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-83-3
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
two ars operons, ars1 and ars2, the arsA gene is split in halves, amarsA1 and amarsA2, and, acr3 but not an arsB gene coexists with arsA
-
-
Manually annotated by BRENDA team
gene asna-1
UniProt
Manually annotated by BRENDA team
gene arsA
-
-
Manually annotated by BRENDA team
plasmid R773
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
gene Rv2643
-
-
Manually annotated by BRENDA team
gene Rv2643
-
-
Manually annotated by BRENDA team
ArsB fragment; genes arsA and arsB
UniProt
Manually annotated by BRENDA team
genes strop634 and strop1447 encoding ACR2 and Ars2, respectively
-
-
Manually annotated by BRENDA team
plasmid pSX267
-
-
Manually annotated by BRENDA team
isolates SPK14, SP45 and SP9
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
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 + H2O
ADP + phosphate
show the reaction diagram
-
-
-
-
?
ATP + H2O + antimonite/in
ADP + phosphate + antimonite/out
show the reaction diagram
ATP + H2O + arsenate/in
ADP + phosphate + arsenate/out
show the reaction diagram
-
in absence of the ArsC protein, MW 16000 Da, the the pump only transports arsenite and antimonite. The ArsC protein alters the specificity of the pump to allow recognition and transport of arsenate
-
?
ATP + H2O + arsenite/in
ADP + phosphate + arsenite/out
show the reaction diagram
ATP + H2O + cisplatin/in
ADP + phosphate + cisplatin/out
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 + antimonite/in
ADP + phosphate + antimonite/out
show the reaction diagram
ATP + H2O + arsenite/in
ADP + phosphate + arsenite/out
show the reaction diagram
ATP + H2O + cisplatin/in
ADP + phosphate + cisplatin/out
show the reaction diagram
O43681
-
-
-
?
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
MgATP2-
-
neccasary for the pump function
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
antimonite
-
inhibits arsenite uptake
-
ATP
-
substrate inhibition
diethyl dicarbonate
-
inhibition could be reversed upon subsequent incubation with hydroxylamine
Trypsin
-
trypsin cleaves the ArsA at Arg290 to produce a 32 kDa A1 fragment that is catalytically inactive and remains stable to trypsin digestion, and a slightly smaller A2 fragment which is digested rapidly. The trypsin digestion pattern is much different when all three ligands, ATP, Sb(III), and Mg2+, are added together, conditions that produce activated catalysis, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
antimonite
-
ArsD
-
arsenite
additional information
-
in vivo, cytosolic As(III) is nearly completely complexed with GSH. GSH greatly increases the rate of binding of As(III) to ArsD, but GSH does not affect the As(III)-stimulated ArsA ATPase activity
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.1
arsenite
-
-
0.021 - 2
ATP
additional information
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.792 - 0.945
ATP
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.082
-
M446W
0.327
-
M446W, stimulated with arsenite
28.87
purified recombinant MBP-fusion enzyme stimulated by 0.5 mM Sb3+
additional information
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
transfer of As(III) from ArsD to ArsA occurs in the presence of MgATP at 23C but not at 4C
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
Methanothermobacter thermautotrophicus (strain ATCC 29096 / DSM 1053 / JCM 10044 / NBRC 100330 / Delta H)
Neosartorya fumigata (strain ATCC MYA-4609 / Af293 / CBS 101355 / FGSC A1100)
Schizosaccharomyces pombe (strain 972 / ATCC 24843)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
heterodimer
the enzyme is composed of ATPase subunit ArsA and arsenic-efflux pump ArsB; the enzyme is composed of ATPase subunit ArsA and arsenic-efflux pump ArsB; the enzyme is composed of ATPase subunit ArsA and arsenic-efflux pump ArsB
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
catalytic subunit of the ATP-dependent arsenite pump is encoded by the Escherichia coli plasmid R773
-
hanging drop vapour diffusion method at 30C, space group P21212 when AATP or AMP-PMP is bound, space group I222 in presence of ADP * A1F3
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ATP and Sb(III) synergistically protect wild type ArsA catalytic subunit from trypsin digestion. Subsequent addition of Mg2+ increases trypsin digestion. Mutant D45N and D45A remain protected by ATP and Sb(III) but lose the Mg2+ effect. D45E exhibits an intermediate Mg2+ response
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography
-
recombinant His6-tagged wild-type and mutant ArsA from Escherichia coli strain JM109 to over 95% homogeneity by nickel affinity chromatography
-
recombinant maltose-binding-protein fusion protein from Escherichia coli by amylose affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
-
expression in Escherichia coli JM109 with a His-tag
-
expression of ArsA and ArsD wild-type and mutants in a Saccharomyces cerevisiae two-hybrid system
-
expression of ArsA and ArsD wild-type and mutants in a Saccharomyces cerevisiae two-hybrid system for interaction analysis, overview
-
expression of His6-tagged wild-type and mutant ArsA in Escherichia coli strain JM109. At high expression level wild-type ArsA is located in the cytosol, mutants D142A, D142E, and D142N are also found predominantly in the cytosol at similar levels as the wild type, but D447A and D447E proteins are found as insoluble aggregates, while only trace amounts of D447N can be observed in the soluble fraction
-
expression of recombinant enzyme with a His-tag
-
expression of the enzyme as maltose-binding-protein fusion protein in Escherichia coli
gene arsA, expression of isolated ArsA ATPase in Escherichia coli strain BL21 (DE3)
-
gene arsA, phylogenetic analysis with detailed analysis of clustering in the superfamily, overview
-
gene arsA, phylogenetic analysis, expression of complete ArsA, partial ArsA2, or chimeric NifH-ArsA2 in Escherichia coli strain JM109, BG1754 (arsAB), BG1757 (arsA2B), and BG1791 (nifH-arsA2B)
gene arsB, DNA and amino acid sequence determination and analysis, phylogenetic analysis; gene arsB, DNA and amino acid sequence determination and analysis, phylogenetic analysis; gene arsB, DNA and amino acid sequence determination and analysis, phylogenetic analysis
genes strop634 and strop1447, expression of enzyme mutants in Escherichia coli strain EPI300 with pCC1FOS-based fosmids
-
recombinant expression of His-tagged wild-type and mutant enzymes
-
the ars operon of the plasmid R773 contains a regulatory gene, arsR, and three structural genes, arsA, arsB, and arsC
-
the arsenical resistance operon from IncN plasmid R46 contains the five genes ArsR, arsD, arsA, arsB and arsC. The chromosomal ars operon contains only arsB, arsC, and arsR
-
transfection of T-289 melanoma cells with either ASNA1 sense or ASNA1 antisense constructs, expression analysis
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C108A
-
alteration of either Cys108 or Cys120 to alanine results in loss of metalloid binding to either pre-mixed or copurified AmArsAs, indicating that the Cys108 of AmArsA1 and Cys120 of AmArsA2 form part of the metalloid binding domain
C120A
-
alteration of either Cys108 or Cys120 to alanine results in loss of metalloid binding to either pre-mixed or copurified AmArsAs, indicating that the Cys108 of AmArsA1 and Cys120 of AmArsA2 form part of the metalloid binding domain
C113A/C422A
C172A
site-directed mutagenesis, an ArsA mutant, the mutant shows reduced affinity for Sb(III) compared to the wild-type enzyme
C172A/H453A
site-directed mutagenesis, the ArsA double mutant exhibits significantly decreased affinity for Sb(III)
D137V
-
inactive
D142A
-
site-directed mutagenesis, the mutant is activated by arsenite and antimonite in a similar amount as the wild-type enzyme
D142E
-
site-directed mutagenesis, the mutant is stronger activated by arsenite and antimonite compared to the wild-type enzyme
D142N
-
site-directed mutagenesis, the mutant is activated by arsenite and antimonite in a similar amount as the wild-type enzyme
D303G
-
highly sensitive towards arsenite
D311G
-
no effect of resistance towards arsenite
D320G
-
no effect of resistance towards arsenite
D447A
-
site-directed mutagenesis, the mutant is less activated by arsenite and antimonite compared to the wild-type enzyme
D447E
-
site-directed mutagenesis, the mutant is less activated by arsenite and antimonite compared to the wild-type enzyme
D447N
-
site-directed mutagenesis, the near complete insolubility of D447N ArsA precludes its purification and biochemical characterization
D45A
-
mutant of ArsA, the catalytic subunit of the pump. Inactive enzyme. ATP and Sb(III) synergistically protect wild type ArsA catalytic subunit from trypsin digestion. Subsequent addition of Mg2+ increases trypsin digestion. Mutant D45N and D45A remain protected by ATP and Sb(III) but lose the Mg2+ effect
D45E
-
approximately 5% of the wild type activity with about a 5fold decrease in affinity for Mg2+. ATP and Sb(III) synergistically protect wild type ArsA catalytic subunit from trypsin digestion. Subsequent addition of Mg2+ increases trypsin digestion, D45E exhibits an lower Mg2+ response
D45N
-
mutant of Ars A, the catalytic subunit of the pump. Inactive enzyme. ATP and Sb(III) synergistically protect wild type ArsA catalytic subunit from trypsin digestion. Subsequent addition of Mg2+ increases trypsin digestion. Mutant D45N and D45A remain protected by ATP and Sb(III) but lose the Mg2+ effect
DELTA1-277
-
same arsenite resistance like wild-type when coexpressed with DELTA280-583 or DELTA324-583
DELTA1-320
-
no arsenite resistance in cells expressing that protein in combination with DELTA280-583 but resistance like wild-type expressing cells when expressed in combination with DELTA324-583
DELTA280-583
-
no arsenite resistance in cells expressing that protein in combination with DELTA1-320 but resistance like wild-type expressing cells when expressed in combination with DELTA1-277
DELTA324-583
-
same arsenite resistance like wild-type when coexpressed with DELTA1-320 or DELTA1-277
F120I
-
inactive
G18D
-
mutation in ArsA protein, loss of resistance to the toxic oxyanions as well as inability to extrude arsenite
G18R
-
mutation in ArsA protein, loss of resistance to the toxic oxyanions as well as inability to extrude arsenite
G18S
-
loss of resistance towards arsenite and no nucleotide-binding in A1 NBD
G20S
-
mutation in ArsA protein, loss of resistance to the toxic oxyanions as well as inability to extrude arsenite
G284S
-
no resistance towards arsenite
H138A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H148A
-
cells with this protein were resistant to arsenite and antimonite like wild-type, 3-fold lowering of the antimonite stimulated ATPase activity compared to wild-type enzyme
H148A/S420A
site-directed mutagenesis, the mutant exhibits a half-maximal stimulation similar to that of the wild-type enzyme
H219A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H327A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H359A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H368A
-
cells with this protein were resistant to arsenite and antimonite like wild-type, 3-fold lowering of the antimonite stimulated ATPase activity compared to wild-type enzyme
H388A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H397A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H453A
-
cells with this protein were resistant to arsenite and antimonite like wild-type, 3-fold lowering of the antimonite stimulated ATPase activity compared to wild-type enzyme
H465A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H477A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H520A
-
cells with this protein were resistant to arsenite and antimonite like wild-type
H558A
-
cells with this protein were resistant to arsenite and antimonite like wild-type, 50% lowering of the antimonite stimulated ATPase activity compared to wild-type enzyme
K16Q
-
site-directed mutagenesis, the mutant ArsA shows 70% of wild-type ATPase activity
K335Q
-
site-directed mutagenesis, the mutant ArsA is inactive. K335Q acquires a closed conformation during metalloid-stimulated catalysis that is different from the open conformation of the wild-type
K340E
-
retains about 10% of wild-type ATPase activity, no antimonite-stimulated ATPase activity
M446W
-
no changes in resistance towards arsenite or in specific activity compared to wild-type
M446WG18S
-
binding of nucleotides like wild-type
Q56R
-
the mutant exhibits significant metalloid-stimulated ATPase activity in vitro
R290K
-
no effect of resistance towards arsenite
R290S
-
decreased resistance towards arsenite
T22I
-
mutation in ArsA protein, loss of resistance to the toxic oxyanions as well as inability to extrude arsenite
K16Q
-
site-directed mutagenesis, the mutant ArsA shows 70% of wild-type ATPase activity
-
K335Q
-
site-directed mutagenesis, the mutant ArsA is inactive. K335Q acquires a closed conformation during metalloid-stimulated catalysis that is different from the open conformation of the wild-type
-
C113A/C422A
-
mutant ArsA has basal ATPase activity similar to that of the wild type but lacks metalloid-stimulated activity
-
additional information
Show AA Sequence (1029 entries)
Please use the Sequence Search for a specific query.