BRENDA - Enzyme Database

Spectroscopic studies of the Salmonella enterica adenosyltransferase enzyme SeCobA: molecular-level insight into the mechanism of substrate Cob(II)alamin activation

Pallares, I.G.; Moore, T.C.; Escalante-Semerena, J.C.; Brunold, T.C.; Biochemistry 53, 7969-7982 (2014)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.5.1.17
expressed in Escherichia coli BL21 cells
Salmonella enterica
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.5.1.17
F91W
with cob(II)alamin the mutant shows catalytic activity diminished relative to that of wild type enzyme, while the activity with cob(I)alamin is largely retained
Salmonella enterica
2.5.1.17
F91W/W93F
in this mutant the yield of 4c cob(II)alamin species is reduced more than 10fold from that achieved by the wild type enzyme
Salmonella enterica
2.5.1.17
F91Y
with cob(II)alamin the mutant shows catalytic activity 5fold increased relative to that of wild type enzyme
Salmonella enterica
2.5.1.17
W93A
the mutation completely abolishes the catalytic activity with cob(II)alamin while modest activity is retained with cob(I)alamin
Salmonella enterica
2.5.1.17
W93F
the mutant shows wild type activity
Salmonella enterica
2.5.1.17
W93H
the mutation inhibits the formation of 4c cob(II)alamin
Salmonella enterica
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.0163
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.0254
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.5.1.17
ATP + cob(I)yrinic acid a,c-diamide
Salmonella enterica
-
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
?
2.5.1.17
ATP + cobinamide
Salmonella enterica
-
triphosphate + adenosylcobinamide
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.5.1.17
Salmonella enterica
P31570
serovar typhimurium LT2
-
Purification (Commentary)
EC Number
Commentary
Organism
2.5.1.17
HisTrap nickel affinity column chromatography
Salmonella enterica
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.5.1.17
ATP + cob(I)alamine
-
737702
Salmonella enterica
triphosphate + adenosylcob(I)alamine
-
-
-
?
2.5.1.17
ATP + cob(I)yrinic acid a,c-diamide
-
737702
Salmonella enterica
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
-
?
2.5.1.17
ATP + cob(II)alamine
-
737702
Salmonella enterica
triphosphate + adenosylcob(II)alamine
-
-
-
?
2.5.1.17
ATP + cob(II)inamide
-
737702
Salmonella enterica
triphosphate + adenosylcob(II)inamide
-
-
-
?
2.5.1.17
ATP + cobinamide
-
737702
Salmonella enterica
triphosphate + adenosylcobinamide
-
-
-
?
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.0067
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.0077
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.5.1.17
expressed in Escherichia coli BL21 cells
Salmonella enterica
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.5.1.17
F91W
with cob(II)alamin the mutant shows catalytic activity diminished relative to that of wild type enzyme, while the activity with cob(I)alamin is largely retained
Salmonella enterica
2.5.1.17
F91W/W93F
in this mutant the yield of 4c cob(II)alamin species is reduced more than 10fold from that achieved by the wild type enzyme
Salmonella enterica
2.5.1.17
F91Y
with cob(II)alamin the mutant shows catalytic activity 5fold increased relative to that of wild type enzyme
Salmonella enterica
2.5.1.17
W93A
the mutation completely abolishes the catalytic activity with cob(II)alamin while modest activity is retained with cob(I)alamin
Salmonella enterica
2.5.1.17
W93F
the mutant shows wild type activity
Salmonella enterica
2.5.1.17
W93H
the mutation inhibits the formation of 4c cob(II)alamin
Salmonella enterica
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.0163
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.0254
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.5.1.17
ATP + cob(I)yrinic acid a,c-diamide
Salmonella enterica
-
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
?
2.5.1.17
ATP + cobinamide
Salmonella enterica
-
triphosphate + adenosylcobinamide
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.5.1.17
HisTrap nickel affinity column chromatography
Salmonella enterica
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.5.1.17
ATP + cob(I)alamine
-
737702
Salmonella enterica
triphosphate + adenosylcob(I)alamine
-
-
-
?
2.5.1.17
ATP + cob(I)yrinic acid a,c-diamide
-
737702
Salmonella enterica
triphosphate + adenosylcob(III)yrinic acid a,c-diamide
-
-
-
?
2.5.1.17
ATP + cob(II)alamine
-
737702
Salmonella enterica
triphosphate + adenosylcob(II)alamine
-
-
-
?
2.5.1.17
ATP + cob(II)inamide
-
737702
Salmonella enterica
triphosphate + adenosylcob(II)inamide
-
-
-
?
2.5.1.17
ATP + cobinamide
-
737702
Salmonella enterica
triphosphate + adenosylcobinamide
-
-
-
?
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.0067
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.0077
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.26
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.47
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
2.5.1.17
0.26
-
ATP
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica
2.5.1.17
0.47
-
cob(II)inamide
wild type enzyme, pH and temperature not specified in the publication
Salmonella enterica