Cloned (Comment) | Organism |
---|---|
overexpressed in Escherichia coli BL21(DE3) | Methanosarcina thermophila |
Protein Variants | Comment | Organism |
---|---|---|
Q43W | site-directed mutagenesis, single mutant | Methanosarcina thermophila |
R241A/Q43W | site-directed mutagenesis, double mutant | Methanosarcina thermophila |
R241K/Q43W | site-directed mutagenesis, double mutant | Methanosarcina thermophila |
R91A/Q43W | site-directed mutagenesis, double mutant | Methanosarcina thermophila |
R91K/Q43W | site-directed mutagenesis, double mutant | Methanosarcina thermophila |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.053 | - |
ATP | Q43W, mutant | Methanosarcina thermophila | |
0.08 | - |
ATP | wild-type | Methanosarcina thermophila | |
0.08 | - |
ATP | R241A/Q43W, double mutant | Methanosarcina thermophila | |
0.098 | - |
ADP | wild-type | Methanosarcina thermophila | |
0.147 | - |
ADP | R91A/Q43W, double mutant | Methanosarcina thermophila | |
0.164 | - |
ADP | R91K/Q43W, double mutant | Methanosarcina thermophila | |
0.173 | - |
ADP | Q43W, mutant | Methanosarcina thermophila | |
0.234 | - |
ATP | R91K/Q43W, double mutant | Methanosarcina thermophila | |
0.239 | - |
ADP | R241A/Q43W, double mutant | Methanosarcina thermophila | |
0.26 | - |
acetyl phosphate | Q43W | Methanosarcina thermophila | |
0.33 | - |
acetyl phosphate | R241/Q43W | Methanosarcina thermophila | |
0.47 | - |
acetyl phosphate | wild-type | Methanosarcina thermophila | |
0.539 | - |
ATP | R91A/Q43W, double mutant | Methanosarcina thermophila | |
0.692 | - |
ADP | R241K/Q43W | Methanosarcina thermophila | |
1.02 | - |
acetyl phosphate | R241K/Q43W | Methanosarcina thermophila | |
1.08 | - |
acetyl phosphate | R91K/Q43W | Methanosarcina thermophila | |
1.3 | - |
ATP | R241K/Q43W, double mutant | Methanosarcina thermophila | |
2.58 | - |
acetyl phosphate | R91A/Q43W | Methanosarcina thermophila | |
2.7 | - |
acetate | wild-type | Methanosarcina thermophila | |
5.8 | - |
acetate | R241A/Q43W, double mutant | Methanosarcina thermophila | |
12.4 | - |
acetate | Q43W, mutant | Methanosarcina thermophila | |
48 | - |
acetate | R91K/Q43W, double mutant | Methanosarcina thermophila | |
179 | - |
acetate | R241K/Q43W, double mutant | Methanosarcina thermophila |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | binding constants of enzyme variants with ADP measured | Methanosarcina thermophila |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate | Methanosarcina thermophila | - |
ADP + acetyl phosphate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanosarcina thermophila | P38502 | - |
- |
Purification (Comment) | Organism |
---|---|
judged by SDS-PAGE | Methanosarcina thermophila |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + acetate = ADP + acetyl phosphate | roles for the arginine residues Arg241 and Arg91 in transition state stabilization for catalysis but not in nucleotide binding determined, experimental evidence for domain motion | Methanosarcina thermophila |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
catalytic mechanism analyzed in wild-type and mutant variants, binding constants for the nucleotide substrates indicate that Arg241 is involved in transition state stabilization and not directly involved in nucleotide recognition or binding, or in the domain closure required for catalysis, binding constants of the nucleotide substrates for Arg91 suggest that this residue has a role in transition state stabilization, evidence for domain motion dependent upon nucleotide ligand binding presented, suggestion that Arg91 is important for closure of domain I onto domain II for catalysis | Methanosarcina thermophila |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + acetate | - |
Methanosarcina thermophila | ADP + acetyl phosphate | - |
r | |
ATP + acetate | catalytic mechanism, roles for the arginine residues Arg241 and Arg91 in transition state stabilization for catalysis but not in nucleotide binding determined, experimental evidence for domain motion | Methanosarcina thermophila | ADP + acetyl phosphate | - |
r |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.4 | - |
ATP | R241K/Q43W, double mutant | Methanosarcina thermophila | |
1.83 | - |
ATP | R241A/Q43W, double mutant | Methanosarcina thermophila | |
2.07 | - |
ATP | R91A/Q43W, double mutant | Methanosarcina thermophila | |
2.63 | - |
ADP | R91A/Q43W, double mutant | Methanosarcina thermophila | |
10.5 | - |
ADP | R241K/Q43W, double mutant | Methanosarcina thermophila | |
11.8 | - |
ATP | R91K/Q43W, double mutant | Methanosarcina thermophila | |
14.6 | - |
ADP | R241A/Q43W, double mutant | Methanosarcina thermophila | |
25.7 | - |
ADP | R91K/Q43W, double mutant | Methanosarcina thermophila | |
699 | - |
ATP | Q43W, mutant | Methanosarcina thermophila | |
913 | - |
ATP | wild-type | Methanosarcina thermophila | |
1255 | - |
ADP | wild-type | Methanosarcina thermophila | |
2650 | - |
ADP | Q43W, mutant | Methanosarcina thermophila |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Methanosarcina thermophila |