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Literature summary for 2.7.2.1 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
Search for more literature for 2.7.2.1

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Methanosarcina thermophila

Protein Variants

Protein Variants Comment Organism
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
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
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
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
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
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
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]

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

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.
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
-
 ?
CTP + acetate 50% of the activity relative to that observed with ATP Methanosarcina thermophila CDP + acetyl phosphate
-
 ?
GTP + acetate 85.2% of the activity relative to that observed with ATP Methanosarcina thermophila GDP + acetyl phosphate
-
 ?
ITP + acetate 80% of the activity relative to that observed with ATP Methanosarcina thermophila IDP + acetyl phosphate
-
 ?
TTP + acetate 64.5% of the activity relative to that observed with ATP Methanosarcina thermophila TDP + acetyl phosphate
-
 ?
UTP + acetate 54.8% of the activity relative to that observed with ATP Methanosarcina thermophila UDP + acetyl phosphate
-
 ?

Synonyms

Synonyms Comment Organism
ACK
-
 Methanosarcina thermophila

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

kcat/KM [mM/s]

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