Cloned (Comment) | Organism |
---|---|
gene eutT, overexpression of His-tagged enzyme in Escherichia coli as insoluble protein | Salmonella enterica |
Protein Variants | Comment | Organism |
---|---|---|
C79A | site-directed mutagenesis of EutT, the mutant shows highly reduced activity compared to the wild-type enzyme | Salmonella enterica |
C80A | site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme | Salmonella enterica |
C83A | site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme | Salmonella enterica |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
diphosphate | about 50% inhibition of EutT | Salmonella enterica | |
additional information | no inhibition of EutT by phosphate | Salmonella enterica | |
Ni2+ | 62% inhibition, single-site binding | Salmonella enterica | |
Triphosphate | about 40% inhibition of EutT | Salmonella enterica | |
Zn2+ | 77% inhibition, single-site binding | Salmonella enterica |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Salmonella enterica | |
0.0041 | - |
cob(I)alamin | pH 7.0, 37°C, recombinant enzyme | Salmonella enterica | |
0.01 | - |
ATP | pH 7.0, 37°C, recombinant enzyme | Salmonella enterica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | 32% stimulation | Salmonella enterica | |
additional information | no stimulation by Ca2+, Mg2+, Na+, and K+ | Salmonella enterica |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
cob(I)alamin + ADP | Salmonella enterica | - |
adenosylcobalamin + diphosphate | - |
? | |
cob(I)alamin + ATP + H2O + H+ | Salmonella enterica | - |
adenosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + CTP + H2O + H+ | Salmonella enterica | - |
cytosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + dATP + H2O + H+ | Salmonella enterica | - |
deoxyadenosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + GTP + H2O + H+ | Salmonella enterica | low activity | guanosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + ITP | Salmonella enterica | low activity | hypoxanthosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + UTP | Salmonella enterica | low activity | uranylcobalamin + diphosphate + phosphate | - |
? | |
additional information | Salmonella enterica | EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Salmonella enterica | - |
gene eutT, eutT is no housekeeping gene in contrast to the other enzyme-encoding gene cobA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged EutT from Escherichia coli to 70% purity | Salmonella enterica |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein | Cys79, Cys80, and Cys83 are important for catalysis | Salmonella enterica | |
2 ATP + 2 cob(II)yrinic acid a,c-diamide + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)yrinic acid a,c-diamide + an oxidized flavoprotein | Cys79, Cys80, and Cys83 are important for catalysis | Salmonella enterica |
Renatured (Comment) | Organism |
---|---|
insoluble recombinant enzyme is lysed under anoxic conditions using CHES/glycine buffer, pH 9.5, followed by resuspension of EutT from the pelleted material with 8 mM CHAPS, and centrifugation | Salmonella enterica |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
nucleotide substrate specificty | Salmonella enterica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
cob(I)alamin + ADP | - |
Salmonella enterica | adenosylcobalamin + diphosphate | - |
? | |
cob(I)alamin + ATP + H2O + H+ | - |
Salmonella enterica | adenosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + ATP + H2O + H+ | FMN and NADH are used to reduce cob(III)alamin to cob(I)alamin, the enzyme shows ATP hydrolyzing activity to adenosine and triphosphate in absence of cob(I)alamin | Salmonella enterica | adenosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + CTP + H2O + H+ | - |
Salmonella enterica | cytosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + dATP + H2O + H+ | - |
Salmonella enterica | deoxyadenosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + GTP + H2O + H+ | low activity | Salmonella enterica | guanosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + ITP | low activity | Salmonella enterica | hypoxanthosylcobalamin + diphosphate + phosphate | - |
? | |
cob(I)alamin + UTP | low activity | Salmonella enterica | uranylcobalamin + diphosphate + phosphate | - |
? | |
additional information | EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview | Salmonella enterica | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ATP:cob(I)alamin adenosyltransferase | - |
Salmonella enterica |
EutT | - |
Salmonella enterica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Salmonella enterica |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
ATP | pH 7.0, 37°C, recombinant enzyme | Salmonella enterica | |
0.06 | - |
cob(I)alamin | pH 7.0, 37°C, recombinant enzyme | Salmonella enterica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Salmonella enterica |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 8.5 | 30% of maximal activity at pH 7.5 | Salmonella enterica |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ADP | about 80% of the activity with ATP | Salmonella enterica | |
ATP | preferred cofactor, the beta-phosphate is required playing a role in nucleotide recognition and/or binding | Salmonella enterica | |
CTP | about 40% activity compared to ATP | Salmonella enterica | |
dATP | about 25% of the activity with ATP | Salmonella enterica | |
GTP | about 5% activity compared to ATP | Salmonella enterica | |
ITP | about 8% activity compared to ATP | Salmonella enterica | |
additional information | no activity with AMP as cofactor | Salmonella enterica | |
UTP | about 10% activity compared to ATP | Salmonella enterica |