BRENDA - Enzyme Database

Spectroscopic characterization of active-site variants of the PduO-type ATP:corrinoid adenosyltransferase from Lactobacillus reuteri: insights into the mechanism of four-coordinate Co(II)corrinoid formation

Park, K.; Mera, P.E.; Escalante-Semerena, J.C.; Brunold, T.C.; Inorg. Chem. 51, 4482-4494 (2012)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
2.5.1.17
gene pduO, recombinant overexpression of His-tagged wild-type and mutant PduOs in Escherichia coli
Lactobacillus reuteri
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.5.1.17
D35N
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F112A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F112H
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F187A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
R128K
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
R132K
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
S159A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
V186A
site-directed mutagenesis
Lactobacillus reuteri
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)alamin
Lactobacillus reuteri
-
triphosphate + adenosylcob(I)alamine
-
-
?
2.5.1.17
ATP + cobinamide
Lactobacillus reuteri
-
triphosphate + adenosylcobinamide
-
-
?
2.5.1.17
additional information
Lactobacillus reuteri
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
?
-
-
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.5.1.17
Lactobacillus reuteri
-
gene pduO
-
Purification (Commentary)
EC Number
Commentary
Organism
2.5.1.17
recombinant His-tagged wild-type and mutant PduOs from Escherichia coli by nickel affinity chromatography to over 99% homogeneity
Lactobacillus reuteri
Reaction
EC Number
Reaction
Commentary
Organism
2.5.1.17
2 ATP + 2 cob(II)alamin + a reduced flavoprotein = 2 triphosphate + 2 adenosylcob(III)alamin + an oxidized flavoprotein
to overcome the thermodynamically challenging Co2+ -> Co1+ reduction, the enzyme drastically weakens the axial ligand-Co2+ bond so as to generate effectively four-coordinate Co2+-corrinoid species, mechanism, overview. The entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+-corrinoids. Large role of the ATP-induced active-site conformational changes with respect to the formation of 4c Co(II)Cbl
Lactobacillus reuteri
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)alamin
-
722386
Lactobacillus reuteri
triphosphate + adenosylcob(I)alamine
-
-
-
?
2.5.1.17
ATP + cobinamide
-
722386
Lactobacillus reuteri
triphosphate + adenosylcobinamide
-
-
-
?
2.5.1.17
additional information
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
722386
Lactobacillus reuteri
?
-
-
-
-
2.5.1.17
additional information
to overcome the thermodynamically challenging Co2+ -> Co1+ reduction, the enzyme drastically weakens the axial ligand-Co2+ bond so as to generate effectively four-coordinate Co2+-corrinoid species, mechanism, overview. The entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+-corrinoids. Large role of the ATP-induced active-site conformational changes with respect to the formation of 4c Co(II)Cbl
722386
Lactobacillus reuteri
?
-
-
-
-
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
2.5.1.17
gene pduO, recombinant overexpression of His-tagged wild-type and mutant PduOs in Escherichia coli
Lactobacillus reuteri
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.5.1.17
D35N
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F112A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F112H
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
F187A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
R128K
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
R132K
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
S159A
site-directed mutagenesis
Lactobacillus reuteri
2.5.1.17
V186A
site-directed mutagenesis
Lactobacillus reuteri
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)alamin
Lactobacillus reuteri
-
triphosphate + adenosylcob(I)alamine
-
-
?
2.5.1.17
ATP + cobinamide
Lactobacillus reuteri
-
triphosphate + adenosylcobinamide
-
-
?
2.5.1.17
additional information
Lactobacillus reuteri
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
?
-
-
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
2.5.1.17
recombinant His-tagged wild-type and mutant PduOs from Escherichia coli by nickel affinity chromatography to over 99% homogeneity
Lactobacillus reuteri
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)alamin
-
722386
Lactobacillus reuteri
triphosphate + adenosylcob(I)alamine
-
-
-
?
2.5.1.17
ATP + cobinamide
-
722386
Lactobacillus reuteri
triphosphate + adenosylcobinamide
-
-
-
?
2.5.1.17
additional information
The PduO-type ATP:corrinoid adenosyltransferase catalyzes the transfer of the adenosyl-group of ATP to Co1+-cobalamin and Co1+-cobinamide substrates to synthesize adenosylcobalamin and adenosylcobinamide, respectively
722386
Lactobacillus reuteri
?
-
-
-
-
2.5.1.17
additional information
to overcome the thermodynamically challenging Co2+ -> Co1+ reduction, the enzyme drastically weakens the axial ligand-Co2+ bond so as to generate effectively four-coordinate Co2+-corrinoid species, mechanism, overview. The entire hydrophobic pocket below the corrin ring, and not just residue F112, is critical for the removal of the axial ligand from the cobalt center of the Co2+-corrinoids. Large role of the ATP-induced active-site conformational changes with respect to the formation of 4c Co(II)Cbl
722386
Lactobacillus reuteri
?
-
-
-
-
General Information
EC Number
General Information
Commentary
Organism
2.5.1.17
additional information
active-site region of wild-type and mutant PduOs complexed with Co(II)Cbl and cosubstrate ATP, structure determination and modeling, overview
Lactobacillus reuteri
General Information (protein specific)
EC Number
General Information
Commentary
Organism
2.5.1.17
additional information
active-site region of wild-type and mutant PduOs complexed with Co(II)Cbl and cosubstrate ATP, structure determination and modeling, overview
Lactobacillus reuteri