Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Buan, N.R.; Escalante-Semerena, J.C.
    Purification and initial biochemical characterization of ATP:Cob(I)alamin adenosyltransferase (EutT) enzyme of Salmonella enterica (2006), J. Biol. Chem., 281, 16971-16977.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.17 gene eutT, overexpression of His-tagged enzyme in Escherichia coli as insoluble protein Salmonella enterica

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.17 C79A site-directed mutagenesis of EutT, the mutant shows highly reduced activity compared to the wild-type enzyme Salmonella enterica
2.5.1.17 C80A site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme Salmonella enterica
2.5.1.17 C83A site-directed mutagenesis of EutT, the mutant shows 99% reduced activity compared to the wild-type enzyme Salmonella enterica

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.17 diphosphate about 50% inhibition of EutT Salmonella enterica
2.5.1.17 additional information no inhibition of EutT by phosphate Salmonella enterica
2.5.1.17 Ni2+ 62% inhibition, single-site binding Salmonella enterica
2.5.1.17 Triphosphate about 40% inhibition of EutT Salmonella enterica
2.5.1.17 Zn2+ 77% inhibition, single-site binding Salmonella enterica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.17 additional information
-
 additional information kinetics Salmonella enterica
2.5.1.17 0.0041
-
 cob(I)alamin pH 7.0, 37°C, recombinant enzyme Salmonella enterica
2.5.1.17 0.01
-
 ATP pH 7.0, 37°C, recombinant enzyme Salmonella enterica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.17 Mn2+ 32% stimulation Salmonella enterica
2.5.1.17 additional information no stimulation by Ca2+, Mg2+, Na+, and K+ Salmonella enterica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.17 cob(I)alamin + ADP Salmonella enterica
-
 adenosylcobalamin + diphosphate
-
 ?
2.5.1.17 cob(I)alamin + ATP + H2O + H+ Salmonella enterica
-
 adenosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + CTP + H2O + H+ Salmonella enterica
-
 cytosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + dATP + H2O + H+ Salmonella enterica
-
 deoxyadenosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + GTP + H2O + H+ Salmonella enterica low activity guanosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + ITP Salmonella enterica low activity hypoxanthosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + UTP Salmonella enterica low activity uranylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 additional information Salmonella enterica EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.17 Salmonella enterica
-
 gene eutT, eutT is no housekeeping gene in contrast to the other enzyme-encoding gene cobA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.17 recombinant His-tagged EutT from Escherichia coli to 70% purity Salmonella enterica

Reaction

EC Number Reaction Comment Organism Reaction ID
2.5.1.17 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
a
2.5.1.17 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
a

Renatured (Commentary)

EC Number Renatured (Comment) Organism
2.5.1.17 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 [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.5.1.17 additional information
-
 nucleotide substrate specificty Salmonella enterica

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.17 cob(I)alamin + ADP
-
 Salmonella enterica adenosylcobalamin + diphosphate
-
 ?
2.5.1.17 cob(I)alamin + ATP + H2O + H+
-
 Salmonella enterica adenosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 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
-
 ?
2.5.1.17 cob(I)alamin + CTP + H2O + H+
-
 Salmonella enterica cytosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + dATP + H2O + H+
-
 Salmonella enterica deoxyadenosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + GTP + H2O + H+ low activity Salmonella enterica guanosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + ITP low activity Salmonella enterica hypoxanthosylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 cob(I)alamin + UTP low activity Salmonella enterica uranylcobalamin + diphosphate + phosphate
-
 ?
2.5.1.17 additional information EuT substrate specificity, overview, reaction cycle involving EutT and CobA, overview Salmonella enterica ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.17 ATP:cob(I)alamin adenosyltransferase
-
 Salmonella enterica
2.5.1.17 EutT
-
 Salmonella enterica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.17 37
-
 assay at Salmonella enterica

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.17 0.03
-
 ATP pH 7.0, 37°C, recombinant enzyme Salmonella enterica
2.5.1.17 0.06
-
 cob(I)alamin pH 7.0, 37°C, recombinant enzyme Salmonella enterica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.17 7
-
 assay at Salmonella enterica

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.5.1.17 7 8.5 30% of maximal activity at pH 7.5 Salmonella enterica

Cofactor

EC Number Cofactor Comment Organism Structure
2.5.1.17 ADP about 80% of the activity with ATP Salmonella enterica
2.5.1.17 ATP preferred cofactor, the beta-phosphate is required playing a role in nucleotide recognition and/or binding Salmonella enterica
2.5.1.17 CTP about 40% activity compared to ATP Salmonella enterica
2.5.1.17 dATP about 25% of the activity with ATP Salmonella enterica
2.5.1.17 GTP about 5% activity compared to ATP Salmonella enterica
2.5.1.17 ITP about 8% activity compared to ATP Salmonella enterica
2.5.1.17 additional information no activity with AMP as cofactor Salmonella enterica
2.5.1.17 UTP about 10% activity compared to ATP Salmonella enterica