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Literature summary extracted from

  • Yoshioka, A.; Murata, K.; Kawai, S.
    Structural and mutational analysis of amino acid residues involved in ATP specificity of Escherichia coli acetate kinase (2014), J. Biosci. Bioeng., 118, 502-507.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.2.1 gene ackA, sequence comparisons, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha Escherichia coli
2.7.2.12 sequence comparisons, recombinant overexpression of His-tagged enzyme in Escherichia coli strain MK3648 Entamoeba histolytica

Protein Variants

EC Number Protein Variants Comment Organism
2.7.2.1 G332D site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli
2.7.2.1 G333Q site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli
2.7.2.1 I334M site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli
2.7.2.1 additional information each of five candidate residue in Escherichia coli ATP-specific AK (ATP-ecoAK), which is unable to use diphosphate, is substituted with the respective diphosphate-ehiAK amino acid residue. Each variant ATP-ecoAK has an increased Km for ATP, indicating that the five residues are the determinants for the specificity to ATP in ATP-ecoAK Escherichia coli
2.7.2.1 N213T site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli
2.7.2.1 N213T/G332D/E336L/T385N site-directed mutagenesis, the mutant shows unaltered Km for ATP and highly reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli
2.7.2.1 N337E site-directed mutagenesis, the mutant shows increased Km for ATP and reduced ATP-dependent acetate kinase activity compared to the wild-type enzyme Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.2.1 additional information
-
 additional information Michaelis-Menten kinetics Escherichia coli
2.7.2.1 0.66
-
 ATP pH 6.5, 30°C, recombinant mutant N213T/G332D/E336L/T385N Escherichia coli
2.7.2.1 0.67
-
 ATP pH 6.5, 30°C, recombinant wild-type enzyme Escherichia coli
2.7.2.1 1.7
-
 ATP pH 6.5, 30°C, recombinant mutant N213T Escherichia coli
2.7.2.1 3.5
-
 ATP pH 6.5, 30°C, recombinant mutant G332D Escherichia coli
2.7.2.1 5.2
-
 ATP pH 6.5, 30°C, recombinant mutant G333Q Escherichia coli
2.7.2.1 6.6
-
 ATP pH 6.5, 30°C, recombinant mutant I334M Escherichia coli
2.7.2.1 30.7
-
 ATP pH 6.5, 30°C, recombinant mutant N337E Escherichia coli
2.7.2.12 additional information
-
 additional information Michaelis-Menten kinetics Entamoeba histolytica
2.7.2.12 3.3
-
 diphosphate pH 6.5-7.5, 30°C, recombinant enzyme Entamoeba histolytica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.2.1 Mg2+ required Escherichia coli
2.7.2.12 Mg2+ required Entamoeba histolytica

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.7.2.12 44000
-
 x * 44000, recombinant His-tagged enzyme, SDS-PAGE Entamoeba histolytica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.2.1 ATP + acetate Escherichia coli
-
 ADP + acetyl phosphate
-
 r
2.7.2.12 diphosphate + acetate Entamoeba histolytica
-
 phosphate + acetyl phosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.7.2.1 Escherichia coli P0A6A3 gene ackA
-
2.7.2.12 Entamoeba histolytica
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.1 recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha by ATP affinity chromatography Escherichia coli

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.2.1 acetyl phosphate + hydroxylamine
-
 Escherichia coli acetyl hydroxamate + phosphate
-
 ?
2.7.2.1 ATP + acetate
-
 Escherichia coli ADP + acetyl phosphate
-
 r
2.7.2.1 additional information no activity with diphosphate Escherichia coli ?
-
 ?
2.7.2.12 diphosphate + acetate
-
 Entamoeba histolytica phosphate + acetyl phosphate
-
 ?
2.7.2.12 additional information no activity with ATP Entamoeba histolytica ?
-
 ?

Subunits

EC Number Subunits Comment Organism
2.7.2.12 ? x * 44000, recombinant His-tagged enzyme, SDS-PAGE Entamoeba histolytica

Synonyms

EC Number Synonyms Comment Organism
2.7.2.1 ackA
-
 Escherichia coli
2.7.2.1 ATP-ecoAK
-
 Escherichia coli
2.7.2.1 ATP-specific AK
-
 Escherichia coli
2.7.2.12 diphosphate-specific AK
-
 Entamoeba histolytica
2.7.2.12 PPi-dependent AK
-
 Entamoeba histolytica
2.7.2.12 PPi-ehiAK
-
 Entamoeba histolytica
2.7.2.12 pyrophosphate-dependent AK
-
 Entamoeba histolytica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.2.1 30
-
 assay at Escherichia coli
2.7.2.12 30
-
 assay at Entamoeba histolytica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.2.1 0.34
-
 ATP pH 6.5, 30°C, recombinant mutant N213T/G332D/E336L/T385N Escherichia coli
2.7.2.1 9.9
-
 ATP pH 6.5, 30°C, recombinant mutant G332D Escherichia coli
2.7.2.1 10.5
-
 ATP pH 6.5, 30°C, recombinant mutant I334M Escherichia coli
2.7.2.1 16.3
-
 ATP pH 6.5, 30°C, recombinant mutant N213T Escherichia coli
2.7.2.1 41
-
 ATP pH 6.5, 30°C, recombinant mutant G333Q Escherichia coli
2.7.2.1 395
-
 ATP pH 6.5, 30°C, recombinant mutant N337E Escherichia coli
2.7.2.1 908
-
 ATP pH 6.5, 30°C, recombinant wild-type enzyme Escherichia coli
2.7.2.12 2.6
-
 diphosphate pH 6.5-7.5, 30°C, recombinant enzyme Entamoeba histolytica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.2.1 6.5 7.5 acetyl phosphate-forming reaction direction, assay at Escherichia coli
2.7.2.12 6.5
-
 assay at, diphosphate forming reaction direction Entamoeba histolytica
2.7.2.12 6.5 7.5 acetyl phosphate-forming reaction direction, assay at Entamoeba histolytica

Cofactor

EC Number Cofactor Comment Organism Structure
2.7.2.1 ADP
-
 Escherichia coli
2.7.2.1 ATP
-
 Escherichia coli
2.7.2.1 additional information no activity with diphosphate as phosphoryl donor Escherichia coli
2.7.2.12 diphosphate purified enzyme PPi-ehiAK utilizes diphosphate, but not ATP, as a phosphoryl donor Entamoeba histolytica
2.7.2.12 additional information no activity with ATP Entamoeba histolytica

General Information

EC Number General Information Comment Organism
2.7.2.1 evolution residue Asn337 of ATP-ecoAK is particularly significant for the specificity to ATP. The 5 residues are highly conserved in 2625 PPi-ehiAK homologsue implying that almost all organisms have ATP-dependent acetate kinase, EC 2.7.2.1, rather than diphosphate-dependent acetate kinase, EC 2.7.2.12 Escherichia coli
2.7.2.1 additional information substrate-binding site structure and comparison with diphosphate-dependent acetate kinase, EC 2.7.2.12, overview Escherichia coli
2.7.2.12 evolution residue Asn337 of ATP-ecoAK is particularly significant for the specificity to ATP. The five residues are highly conserved in 2625 PPi-ehiAK homologue implying that almost all organisms have ATP-dependent acetate kinase, EC 2.7.2.1, rather than diphosphate-dependent acetate kinase, EC 2.7.2.12 Entamoeba histolytica
2.7.2.12 additional information substrate-binding site structure and comparison with ATP-dependent acetate kinase, EC 2.7.2.1, overview Entamoeba histolytica