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Literature summary for 2.5.1.17 extracted from

  • Buan, N.R.; Escalante-Semerena, J.C.
    Computer-assisted docking of flavodoxin with the ATP:Co(I)rrinoid adenosyltransferase (CobA) enzyme reveals residues critical for protein-protein interactions but not for catalysis (2005), J. Biol. Chem., 280, 40948-40956.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutant enzymes in Escherichia coli using strain BL21(DE3) for the His-tagged variant, and several derivatives of strain DH5alpha, overview Salmonella enterica

Protein Variants

Protein Variants Comment Organism
A134L site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Salmonella enterica
additional information truncation of the 9-amino acid N-terminal helix of CobA reduces its FldA-dependent cobalamin adenosyltransferase activity by 97.4%, but shows 4fold higher specific activity than the wild-type enzyme when cob(I)alamin is generated chemically in situ, e.g. by use of KBH4, residues Arg9 and Arg165 of CobA are critical for FldA-dependent adenosylation but are catalytically as competent as the wild-type protein when cob(I)alamin i provided as substrate, overview Salmonella enterica
R165A site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Salmonella enterica
R165E site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme Salmonella enterica
R98A site-directed mutagenesis, the mutant shows 80% reduced activity compared to the wild-type enzyme Salmonella enterica
R98E site-directed mutagenesis, the mutant shows about 30% reduced activity compared to the wild-type enzyme Salmonella enterica
R98E/R165E site-directed mutagenesis, the mutant shows 85% reduced activity compared to the wild-type enzyme Salmonella enterica
R9A site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme Salmonella enterica
R9E site-directed mutagenesis, the mutant shows slightly increased activity compared to the wild-type enzyme Salmonella enterica
R9E/R165E site-directed mutagenesis, the mutant shows 30% reduced activity compared to the wild-type enzyme Salmonella enterica
R9E/R98E site-directed mutagenesis, the mutant shows 60% reduced activity compared to the wild-type enzyme Salmonella enterica
R9E/R98E/R165E site-directed mutagenesis, the mutant shows 25% reduced activity compared to the wild-type enzyme Salmonella enterica

Inhibitors

Inhibitors Comment Organism Structure
TWYYGEAQCDWDD68 competitive 30% inhibition of flavodoxin-dependent adenosylcobalamin synthesis by a FldA peptide TWYYGEAQCDWDD68, the inhibition depends on residues Glu61 and Asp68, overview, no inhibition by peptides TWYYGAAQCDWDA68 and WPTAGYHFEASKG132 Salmonella enterica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information
-
Salmonella enterica

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+
-
Salmonella enterica

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
cob(I)alamin + ATP Salmonella enterica the enzyme is required to adenosylate de novo biosynthetic intermediates of adenosylcobalamin and to salvage incomplete and complete corrinoids from the environment of this bacterium adenosylcobalamin + triphosphate
-
?

Organism

Organism UniProt Comment Textmining
Salmonella enterica
-
sv. typhimurium, several strains, overview, gene cobA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) 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 reaction mechanism 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 reaction mechanism Salmonella enterica

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
activity of mutant enzymes, overview Salmonella enterica
0.143
-
purified recombinant wild-type enzyme Salmonella enterica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
cob(I)alamin + ATP the enzyme is required to adenosylate de novo biosynthetic intermediates of adenosylcobalamin and to salvage incomplete and complete corrinoids from the environment of this bacterium Salmonella enterica adenosylcobalamin + triphosphate
-
?
cob(I)alamin + ATP in vitro reduced flavodoxin provides an electron to generate the co(I)rrinoid substrate in the CobA active site, modeling of enzyme ligand interaction, residues R9 and R165 are important for CobA-FldA docking but not to catalysis, overview Salmonella enterica adenosylcobalamin + triphosphate
-
?

Synonyms

Synonyms Comment Organism
ATP:co(I)rrinoid adenosyltransferase
-
Salmonella enterica
CobA
-
Salmonella enterica

Temperature Optimum [°C]

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

Cofactor

Cofactor Comment Organism Structure
ATP
-
Salmonella enterica
flavodoxin A in vitro reduced flavodoxin provides an electron to generate the co(I)rrinoid substrate in the CobA active site, modeling of enzyme ligand interaction, residues R9 and R165 are important for CobA-FldA docking but not to catalysis, overview Salmonella enterica

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0013
-
TWYYGEAQCDWDD68 37°C, recombinant wild-type enzyme Salmonella enterica