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Literature summary for 2.7.2.1 extracted from

  • Dang, T.; Ingram-Smith, C.
    Investigation of pyrophosphate versus ATP substrate selection in the Entamoeba histolytica acetate kinase (2017), Sci. Rep., 7, 5912 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 2.7.2.1

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta2 (DE3) pLysS cells Methanosarcina thermophila

Protein Variants

Protein Variants Comment Organism
G331Q/I332M the mutations result in substantial reductions in kcat compared to the wild type enzyme. In the acetate-forming direction, catalysis is reduced over 100fold, and in the acetyl phosphate-forming direction, kcat is reduced about 50fold. This alteration results in about 5fold increase in Km for ADP and ATP, and a 15fold increase in Km for acetate but no substantial change in the Km for acetyl phosphate Methanosarcina thermophila

Inhibitors

Inhibitors Comment Organism Structure
diphosphate about 70% inhibition in the acetate-forming direction and about 90% inhibition in the acetyl phosphate-forming direction Methanosarcina thermophila
additional information not inhibited by phosphate Methanosarcina thermophila

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.2
-
 acetyl phosphate wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila
1.7
-
 acetyl phosphate mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
1.7
-
 ATP wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
1.8
-
 ADP wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila
5.7
-
 ADP mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
9.4
-
 ATP mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila
20
-
 acetate wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
302
-
 acetate mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ 5 mM used in assay conditions Methanosarcina thermophila

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ADP + acetyl phosphate Methanosarcina thermophila
-
 ATP + acetate
-
 r
ATP + acetate Methanosarcina thermophila
-
 ADP + acetyl phosphate
-
 r

Organism

Organism UniProt Comment Textmining
Methanosarcina thermophila P38502
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Methanosarcina thermophila

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ADP + acetyl phosphate
-
 Methanosarcina thermophila ATP + acetate
-
 r
ATP + acetate
-
 Methanosarcina thermophila ADP + acetyl phosphate
-
 r

Synonyms

Synonyms Comment Organism
ACK
-
 Methanosarcina thermophila

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
12
-
 ADP mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
12
-
 acetyl phosphate mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
14
-
 ATP mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila
17
-
 acetate mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila
711
-
 ATP wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
789
-
 acetate wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
1550
-
 acetyl phosphate wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila
1915
-
 ADP wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.3
-
 wild type enzyme, in the acetate-forming direction, at pH 7.0 and 37°C Methanosarcina thermophila diphosphate
4
-
 mutant enzyme G331Q/I332M, in the acetate-forming direction, at pH 7.0 and 37°C Methanosarcina thermophila diphosphate
7.5
-
 wild type enzyme, in the acetyl phosphate-forming direction, at pH 5.5 and 45°C Methanosarcina thermophila diphosphate
14
-
 mutant enzyme G331Q/I332M, in the acetyl phosphate-forming direction, at pH 5.5 and 45°C Methanosarcina thermophila diphosphate

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.057
-
 acetate mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila
1.5
-
 ATP mutant enzyme G331Q/I332M, at pH 5.5 and 45°C Methanosarcina thermophila
2
-
 ADP mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
7.2
-
 acetyl phosphate mutant enzyme G331Q/I332M, at pH 7.0 and 37°C Methanosarcina thermophila
39
-
 acetate wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
421
-
 ATP wild type enzyme, at pH 5.5 and 45°C Methanosarcina thermophila
1073
-
 ADP wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila
1294
-
 acetyl phosphate wild type enzyme, at pH 7.0 and 37°C Methanosarcina thermophila