Activating Compound | Comment | Organism | Structure |
---|---|---|---|
KCl | below 50 mM | Thermoplasma acidophilum |
Cloned (Comment) | Organism |
---|---|
- |
Thermoplasma acidophilum |
expression in Escherichia coli in strain BL21(DE3) of the wild-type enzyme and in strain B834(DE3) as selenomethionine-labeled enzyme | Thermoplasma acidophilum |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type and selenomethionine-labeled enzymes, crystal growth from 0.4 M ammonium phosphate, 4% methyl-pentanediol, 5% glycerol, at 20°C, X-ray diffraction structure determination and analysis at 1.5-1.9 A resolution | Thermoplasma acidophilum |
Protein Variants | Comment | Organism |
---|---|---|
E126A | inactive mutant | Thermoplasma acidophilum |
E126A | no catalytic activity | Thermoplasma acidophilum |
E126A | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
E126K | inactive mutant | Thermoplasma acidophilum |
E126K | no catalytic activity | Thermoplasma acidophilum |
E126K | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
R119A | inactive mutant | Thermoplasma acidophilum |
R119A | no catalytic activity | Thermoplasma acidophilum |
R119A | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
R119W | inactive mutant | Thermoplasma acidophilum |
R119W | no catalytic activity | Thermoplasma acidophilum |
R119W | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
R124A | reduced activity | Thermoplasma acidophilum |
R124A | about half as active as wild-type | Thermoplasma acidophilum |
R124A | site-directed mutagenesis, about 40% reduced activity compared to the wild-type enzyme | Thermoplasma acidophilum |
R124F | reduced activity | Thermoplasma acidophilum |
R124F | decreased kcat and increased KM | Thermoplasma acidophilum |
R124F | site-directed mutagenesis, about 95% reduced activity compared to the wild-type enzyme | Thermoplasma acidophilum |
R124K | inactive mutant | Thermoplasma acidophilum |
R124K | no catalytic activity | Thermoplasma acidophilum |
R124K | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
R124W | inactive mutant | Thermoplasma acidophilum |
R124W | no catalytic activity | Thermoplasma acidophilum |
R124W | site-directed mutagenesis, inactive mutant | Thermoplasma acidophilum |
General Stability | Organism |
---|---|
2-mercaptoethanol stabilizes the enzyme during purfication | Thermoplasma acidophilum |
stable | Thermoplasma acidophilum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
hydroxocobalamin | - |
Thermoplasma acidophilum | |
0.003 | - |
hydroxocobalamin | +/- 0.0004 | Thermoplasma acidophilum | |
0.003 | - |
Cobalamin | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum | |
0.004 | - |
Cobalamin | pH 8.0, 70°C, mutant R124A, in presence of Mg2+ | Thermoplasma acidophilum | |
0.11 | - |
ATP | - |
Thermoplasma acidophilum | |
0.11 | - |
Mg2+ | pH 8.0, 70°C, wild-type enzyme | Thermoplasma acidophilum | |
0.11 | - |
Mn2+ | pH 8.0, 70°C, wild-type enzyme | Thermoplasma acidophilum | |
0.11 | - |
ATP | +/- 0.01 | Thermoplasma acidophilum | |
0.11 | - |
ATP | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum | |
0.14 | - |
dATP | - |
Thermoplasma acidophilum | |
0.14 | - |
dATP | +/- 0.02 | Thermoplasma acidophilum | |
0.14 | - |
dATP | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum | |
0.27 | - |
Co2+ | pH 8.0, 70°C, wild-type enzyme | Thermoplasma acidophilum | |
0.33 | - |
ATP | pH 8.0, 70°C, mutant R124A, in presence of Mg2+ | Thermoplasma acidophilum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | activates, KD: 0.27 mM | Thermoplasma acidophilum | |
Co2+ | enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+, 39% of the activity with Mg2+ | Thermoplasma acidophilum | |
Mg2+ | activates, KD: 0.11 mM | Thermoplasma acidophilum | |
Mg2+ | enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+ | Thermoplasma acidophilum | |
Mn2+ | activates, KD: 0.11 mM | Thermoplasma acidophilum | |
Mn2+ | enzyme depends absolutely on divalent cations in the descending order Mg2+, Mn2+, Co2+, 83% of the activity with Mg2+ | Thermoplasma acidophilum | |
additional information | no activity with Ni2+ and Ca2+ | Thermoplasma acidophilum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + cob(I)alamin | Thermoplasma acidophilum | - |
triphosphate + adenosylcobalamin | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermoplasma acidophilum | Q9HIA7 | - |
- |
Thermoplasma acidophilum | Q9HIA7 | gene TA1434 | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.034 | - |
recombinant mutant R124F | Thermoplasma acidophilum |
0.28 | - |
dATP | Thermoplasma acidophilum |
0.28 | - |
+/- 0.01, dATP | Thermoplasma acidophilum |
0.385 | - |
recombinant mutant R124A | Thermoplasma acidophilum |
0.47 | - |
hydroxocobalamin | Thermoplasma acidophilum |
0.47 | - |
+/- 0.02, hydroxocobalamin | Thermoplasma acidophilum |
0.61 | - |
ATP | Thermoplasma acidophilum |
0.61 | - |
+/- 0.02, ATP | Thermoplasma acidophilum |
0.619 | - |
recombinant wild-type enzyme | Thermoplasma acidophilum |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + cob(I)alamin | - |
Thermoplasma acidophilum | triphosphate + adenosylcobalamin | - |
? | |
ATP + cobalamin | enzyme is absolutely specific for ATP or dATP as adenosyl donors | Thermoplasma acidophilum | triphosphate + adenosylcobalamin | - |
ir | |
ATP + hydroxocobalamin | - |
Thermoplasma acidophilum | ? | - |
? | |
dATP + cob(I)alamin | - |
Thermoplasma acidophilum | tripolyphosphate + deoxyadenosylcobalamin | - |
? | |
dATP + cobalamin | enzyme is absolutely specific for ATP or dATP as adenosyl donors | Thermoplasma acidophilum | triphosphate + deoxyadenosylcobalamin | - |
ir | |
additional information | conserved residues are R119, R124, and E126 | Thermoplasma acidophilum | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | each subunit is composed of a bundle of 5 alpha-helices, the active site lies at the junction between the subunits | Thermoplasma acidophilum |
trimer | - |
Thermoplasma acidophilum |
trimer | gel filtration | Thermoplasma acidophilum |
trimer | gel-filtration | Thermoplasma acidophilum |
Synonyms | Comment | Organism |
---|---|---|
ATP:cobalamin adenosyltransferase | - |
Thermoplasma acidophilum |
MMAB | - |
Thermoplasma acidophilum |
TA1434 | - |
Thermoplasma acidophilum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
- |
Thermoplasma acidophilum |
70 | - |
assay at | Thermoplasma acidophilum |
Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | 80 | - |
Thermoplasma acidophilum |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
dATP | - |
Thermoplasma acidophilum | |
0.11 | - |
dATP | +/- 0.01 | Thermoplasma acidophilum | |
0.11 | - |
Cobalamin | pH 8.0, 70°C, mutant R124A, in presence of Mg2+ | Thermoplasma acidophilum | |
0.11 | - |
dATP | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum | |
0.14 | - |
ATP | pH 8.0, 70°C, mutant R124A, in presence of Mg2+ | Thermoplasma acidophilum | |
0.18 | - |
hydroxocobalamin | - |
Thermoplasma acidophilum | |
0.18 | - |
hydroxocobalamin | +/- 0.01 | Thermoplasma acidophilum | |
0.18 | - |
Cobalamin | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum | |
0.23 | - |
ATP | - |
Thermoplasma acidophilum | |
0.23 | - |
ATP | +/- 0.01 | Thermoplasma acidophilum | |
0.23 | - |
ATP | pH 8.0, 70°C, wild-type enzyme, in presence of Mg2+ | Thermoplasma acidophilum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Thermoplasma acidophilum |
8 | - |
assay at | Thermoplasma acidophilum |