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

BRENDA support

Literature summary for 2.5.1.17 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)

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

Protein Variants

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

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 [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

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

Natural Substrates/ Products (Substrates)

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

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

Purification (Commentary)

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

Reaction

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 (Commentary)

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 [micromol/min/mg]

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

Substrates and Products (Substrate)

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

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

Temperature Optimum [°C]

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

Turnover Number [1/s]

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

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

pH Range

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

Cofactor

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