BRENDA - Enzyme Database show

Role of signature lysines in the deviant walker a motifs of the ArsA ATPase

Fu, H.; Ajees, A.; Rosen, B.; Bhattacharjee, H.; Biochemistry 49, 356-364 (2010)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
7.3.2.7
recombinant expression of His-tagged wild-type and mutant enzymes
Escherichia coli
Engineering
EC Number
Amino acid exchange
Commentary
Organism
7.3.2.7
K16Q
site-directed mutagenesis, the mutant ArsA shows 70% of wild-type ATPase activity
Escherichia coli
7.3.2.7
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
Escherichia coli
Metals/Ions
EC Number
Metals/Ions
Commentary
Organism
Structure
7.3.2.7
Mg2+
required
Escherichia coli
7.3.2.7
Sb3+
ArsA is activated by binding of Sb(III), and both wild-type and mutant K16Q ArsAs bind Sb(III) with a 1:1 stoichiometry
Escherichia coli
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
7.3.2.7
63000
-
x * 63000, recombinant ArsA, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7.3.2.7
ATP + H2O + antimonite/in
Escherichia coli
-
ADP + phosphate + antimonite/out
-
-
-
7.3.2.7
ATP + H2O + antimonite/in
Escherichia coli JM109
-
ADP + phosphate + antimonite/out
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
Escherichia coli
-
ADP + phosphate + arsenite/out
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
Escherichia coli JM109
-
ADP + phosphate + arsenite/out
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
7.3.2.7
Escherichia coli
-
gene arsA
-
7.3.2.7
Escherichia coli JM109
-
gene arsA
-
Purification (Commentary)
EC Number
Commentary
Organism
7.3.2.7
recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography
Escherichia coli
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7.3.2.7
ATP + H2O + antimonite/in
-
718867
Escherichia coli
ADP + phosphate + antimonite/out
-
-
-
-
7.3.2.7
ATP + H2O + antimonite/in
-
718867
Escherichia coli JM109
ADP + phosphate + antimonite/out
-
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
-
718867
Escherichia coli
ADP + phosphate + arsenite/out
-
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
-
718867
Escherichia coli JM109
ADP + phosphate + arsenite/out
-
-
-
-
7.3.2.7
additional information
Lys16 is not critical for ATPase activity, while Lys335 is involved in intersubunit interaction and activation of ATPase activity in both halves of the protein
718867
Escherichia coli
?
-
-
-
-
7.3.2.7
additional information
Lys16 is not critical for ATPase activity, while Lys335 is involved in intersubunit interaction and activation of ATPase activity in both halves of the protein
718867
Escherichia coli JM109
?
-
-
-
-
Subunits
EC Number
Subunits
Commentary
Organism
7.3.2.7
?
x * 63000, recombinant ArsA, SDS-PAGE
Escherichia coli
7.3.2.7
More
ArsA structure analysis and comparison, PDB entry 1F48, overview
Escherichia coli
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
7.3.2.7
recombinant expression of His-tagged wild-type and mutant enzymes
Escherichia coli
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
7.3.2.7
K16Q
site-directed mutagenesis, the mutant ArsA shows 70% of wild-type ATPase activity
Escherichia coli
7.3.2.7
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
Escherichia coli
Metals/Ions (protein specific)
EC Number
Metals/Ions
Commentary
Organism
Structure
7.3.2.7
Mg2+
required
Escherichia coli
7.3.2.7
Sb3+
ArsA is activated by binding of Sb(III), and both wild-type and mutant K16Q ArsAs bind Sb(III) with a 1:1 stoichiometry
Escherichia coli
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
7.3.2.7
63000
-
x * 63000, recombinant ArsA, SDS-PAGE
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
7.3.2.7
ATP + H2O + antimonite/in
Escherichia coli
-
ADP + phosphate + antimonite/out
-
-
-
7.3.2.7
ATP + H2O + antimonite/in
Escherichia coli JM109
-
ADP + phosphate + antimonite/out
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
Escherichia coli
-
ADP + phosphate + arsenite/out
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
Escherichia coli JM109
-
ADP + phosphate + arsenite/out
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
7.3.2.7
recombinant His-tagged wild-type and mutant enzymes by nickel affinity chromatography
Escherichia coli
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
7.3.2.7
ATP + H2O + antimonite/in
-
718867
Escherichia coli
ADP + phosphate + antimonite/out
-
-
-
-
7.3.2.7
ATP + H2O + antimonite/in
-
718867
Escherichia coli JM109
ADP + phosphate + antimonite/out
-
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
-
718867
Escherichia coli
ADP + phosphate + arsenite/out
-
-
-
-
7.3.2.7
ATP + H2O + arsenite/in
-
718867
Escherichia coli JM109
ADP + phosphate + arsenite/out
-
-
-
-
7.3.2.7
additional information
Lys16 is not critical for ATPase activity, while Lys335 is involved in intersubunit interaction and activation of ATPase activity in both halves of the protein
718867
Escherichia coli
?
-
-
-
-
7.3.2.7
additional information
Lys16 is not critical for ATPase activity, while Lys335 is involved in intersubunit interaction and activation of ATPase activity in both halves of the protein
718867
Escherichia coli JM109
?
-
-
-
-
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
7.3.2.7
?
x * 63000, recombinant ArsA, SDS-PAGE
Escherichia coli
7.3.2.7
More
ArsA structure analysis and comparison, PDB entry 1F48, overview
Escherichia coli
General Information
EC Number
General Information
Commentary
Organism
7.3.2.7
evolution
the ArsA ATPase belongs to the P-loop GTPase subgroup within the GTPase superfamily of proteins, members of this subgroup have a deviant Walker A motif
Escherichia coli
7.3.2.7
malfunction
while Lys16 mutants show similar resistance phenotypes as the wild type, the Lys335 mutants are sensitive to higher concentrations of arsenite. The As(III)/Sb(III) binding affinity decreases in the order ArsA wild-type > K16Q > K335Q
Escherichia coli
7.3.2.7
additional information
ARsA ATPase contains a deviant Walker A motif which has a signature lysine that is predicted to make intermonomer contact with the bound nucleotides and to play a role in ATP hydrolysis. ArsA has two signature lysines located at positions 16 and 335. Both wild-type and K16Q adopt a similar conformation during activated catalysis, whereas K335Q adopts a conformation that is resistant to trypsin cleavage
Escherichia coli
7.3.2.7
physiological function
the ArsAB pump in Escherichia coli, encoded by the ars operon of plasmid R773, confers resistance to arsenicals and antimonials. ArsA is the catalytic subunit of the pump that hydrolyzesATP in the presence of arsenite [As(III)] or antimonite [Sb(III)]. ATP hydrolysis is coupled to extrusion of As(III) or Sb(III) through ArsB, which serves both as a membrane anchor for ArsA and as the substrate-conducting pathway
Escherichia coli
General Information (protein specific)
EC Number
General Information
Commentary
Organism
7.3.2.7
evolution
the ArsA ATPase belongs to the P-loop GTPase subgroup within the GTPase superfamily of proteins, members of this subgroup have a deviant Walker A motif
Escherichia coli
7.3.2.7
malfunction
while Lys16 mutants show similar resistance phenotypes as the wild type, the Lys335 mutants are sensitive to higher concentrations of arsenite. The As(III)/Sb(III) binding affinity decreases in the order ArsA wild-type > K16Q > K335Q
Escherichia coli
7.3.2.7
additional information
ARsA ATPase contains a deviant Walker A motif which has a signature lysine that is predicted to make intermonomer contact with the bound nucleotides and to play a role in ATP hydrolysis. ArsA has two signature lysines located at positions 16 and 335. Both wild-type and K16Q adopt a similar conformation during activated catalysis, whereas K335Q adopts a conformation that is resistant to trypsin cleavage
Escherichia coli
7.3.2.7
physiological function
the ArsAB pump in Escherichia coli, encoded by the ars operon of plasmid R773, confers resistance to arsenicals and antimonials. ArsA is the catalytic subunit of the pump that hydrolyzesATP in the presence of arsenite [As(III)] or antimonite [Sb(III)]. ATP hydrolysis is coupled to extrusion of As(III) or Sb(III) through ArsB, which serves both as a membrane anchor for ArsA and as the substrate-conducting pathway
Escherichia coli