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Literature summary extracted from

  • Ingram-Smith, C.; Wharton, J.; Reinholz, C.; Doucet, T.; Hesler, R.; Smith, K.
    The role of active site residues in ATP binding and catalysis in the Methanosarcina thermophila acetate kinase (2015), Life, 5, 861-871.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.2.1 expression in Escherichia coli Methanosarcina thermophila

Protein Variants

EC Number Protein Variants Comment Organism
2.7.2.1 G239A 14.5fold reduced specific activity (with ATP as substrate). The wild type enzyme shows broad NTP utilization, with greater than 50% activity with CTP, GTP, TTP, UTP, and ITP versus ATP. The mutant enzyme shows a shift in NTP utilization. It displays substantially higher activity with TTP than with ATP, and activity (as a percentage of that observed with ATP) increased greatly with CTP as well. A weak increase in activity is observed with UTP. Activity with the purines GTP and ITP versus ATP decreases Methanosarcina thermophila
2.7.2.1 G239S 192fold reduced specific activity (with ATP as substrate). The wild type enzyme shows broad NTP utilization, with greater than 50% activity with CTP, GTP, TTP, UTP, and ITP versus ATP. The mutant enzyme shows a shift in NTP utilization. It displays substantially higher activity with TTP than with ATP, and activity (as a percentage of that observed with ATP) increased greatly with CTP as well. A weak increase in activity is observed with UTP. Activity with the purines GTP and ITP versus ATP decreases Methanosarcina thermophila
2.7.2.1 G331A 15.2fold reduced specific activity (with ATP as substrate). The wild type enzyme shows broad NTP utilization, with greater than 50% activity with CTP, GTP, TTP, UTP, and ITP versus ATP. Activity of mutant enzyme with the pyrimidine nucleotides CTP, TTP, and UTP is significantly reduced, with each displaying less than 12% activity versus ATP. Activity with GTP and ITP is also reduced, but to a much lesser extent Methanosarcina thermophila
2.7.2.1 G331Q 118fold reduced specific activity. The wild type enzyme shows broad NTP utilization, with greater than 50% activity with CTP, GTP, TTP, UTP, and ITP versus ATP. Activity of mutant enzyme with the pyrimidine nucleotides CTP, TTP, and UTP is significantly reduced, with each displaying less than 12% activity versus ATP. Activity with GTP and ITP is also reduced, but to a much lesser extent Methanosarcina thermophila
2.7.2.1 N211A 6fold reduced specific activity (with ATP as substrate). The percentage activity observed with CTP and ITP versus ATP is similar to that observed with the wild type enzyme. Activity with GTP and UTP decreases somewhat, and activity with TTP shows an increase Methanosarcina thermophila
2.7.2.1 N211S 200fold reduced specific activity (with ATP as substrate). The percentage activity observed with CTP and ITP versus ATP is similar to that observed with the wild type enzyme. Activity with GTP and UTP decreases somewhat, and activity with TTP shows an increase Methanosarcina thermophila
2.7.2.1 N211T 44fold reduced specific activity (with ATP as substrate). Mutant enzyme shows little change in percentage activity observed with CTP and ITP. Activity with TTP is greatly enhanced and nearly equal to that observed with ATP, whereas the reduction in activity with UTP is stronger than that observed with the N211A Methanosarcina thermophila

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.2.1 1.7
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239A Methanosarcina thermophila
2.7.2.1 1.8
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239S Methanosarcina thermophila
2.7.2.1 2.7
-
UTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 2.7
-
TTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 3.2
-
CTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 4
-
ATP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 4.7
-
ITP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 5.3
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 5.9
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211A Methanosarcina thermophila
2.7.2.1 6.8
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211S Methanosarcina thermophila
2.7.2.1 7.1
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211T Methanosarcina thermophila
2.7.2.1 7.2
-
GTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 7.4
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 10.1
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 10.2
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 10.7
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 11.1
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 11.2
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 12.1
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 14.2
-
UTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 14.3
-
acetate 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 15.7
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 18.4
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 94
-
acetate 37°C, pH 7.0-7.5, mutant enzyme G239S Methanosarcina thermophila
2.7.2.1 113
-
acetate 37°C, pH 7.0-7.5, mutant enzyme G239A Methanosarcina thermophila
2.7.2.1 140
-
acetate 37°C, pH 7.0-7.5, mutant enzyme N211S Methanosarcina thermophila
2.7.2.1 165
-
acetate 37°C, pH 7.0-7.5, mutant enzyme N211A Methanosarcina thermophila
2.7.2.1 470
-
acetate 37°C, pH 7.0-7.5, mutant enzyme N211T Methanosarcina thermophila
2.7.2.1 601
-
acetate 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila

Organism

EC Number Organism UniProt Comment Textmining
2.7.2.1 Methanosarcina thermophila P38502
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.2.1
-
Methanosarcina thermophila

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.2.1 ATP + acetate the wild type enzyme shows broad NTP utilization, with greater than 50% activity with CTP, GTP, TTP, UTP, and ITP versus ATP Methanosarcina thermophila ADP + acetyl phosphate
-
?
2.7.2.1 CTP + acetate 50% of the activity relative to that observed with ATP Methanosarcina thermophila CDP + acetyl phosphate
-
?
2.7.2.1 GTP + acetate 85.2% of the activity relative to that observed with ATP Methanosarcina thermophila GDP + acetyl phosphate
-
?
2.7.2.1 ITP + acetate 80% of the activity relative to that observed with ATP Methanosarcina thermophila IDP + acetyl phosphate
-
?
2.7.2.1 TTP + acetate 64.5% of the activity relative to that observed with ATP Methanosarcina thermophila TDP + acetyl phosphate
-
?
2.7.2.1 UTP + acetate 54.8% of the activity relative to that observed with ATP Methanosarcina thermophila UDP + acetyl phosphate
-
?

Synonyms

EC Number Synonyms Comment Organism
2.7.2.1 ACK
-
Methanosarcina thermophila

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.2.1 37
-
assay at Methanosarcina thermophila
2.7.2.1 65
-
-
Methanosarcina thermophila

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.2.1 2.1
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 2.8
-
UTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 2.8
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 6.8
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239S Methanosarcina thermophila
2.7.2.1 8.6
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 11
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 13
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211S Methanosarcina thermophila
2.7.2.1 16
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 17
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 20
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 32
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211T Methanosarcina thermophila
2.7.2.1 43
-
UTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 67
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 77
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239A Methanosarcina thermophila
2.7.2.1 84
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 88
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 374
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211A Methanosarcina thermophila
2.7.2.1 415
-
UTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 460
-
CTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 540
-
TTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 571
-
GTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 715
-
ATP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 742
-
ITP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7.2.1 0.2
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 0.2
-
UTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 0.2
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 0.9
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 1
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 1.1
-
CTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 1.2
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331Q Methanosarcina thermophila
2.7.2.1 1.8
-
TTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 1.9
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211S Methanosarcina thermophila
2.7.2.1 3.9
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239S Methanosarcina thermophila
2.7.2.1 4.5
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211T Methanosarcina thermophila
2.7.2.1 7.8
-
ITP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 8.9
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 12.6
-
GTP 37°C, pH 7.0-7.5, mutant enzyme G331A Methanosarcina thermophila
2.7.2.1 45.7
-
ATP 37°C, pH 7.0-7.5, mutant enzyme G239A Methanosarcina thermophila
2.7.2.1 63
-
ATP 37°C, pH 7.0-7.5, mutant enzyme N211A Methanosarcina thermophila
2.7.2.1 80
-
GTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 148
-
CTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 152
-
UTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 158
-
ITP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 180
-
ATP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila
2.7.2.1 202
-
TTP 37°C, pH 7.0-7.5, wild-type enzyme Methanosarcina thermophila