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 | 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 | 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 Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Salmonella enterica |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mn2+ | - |
Salmonella enterica |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Salmonella enterica | - |
sv. typhimurium, several strains, overview, gene cobA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) | Salmonella enterica |
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 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 | 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 | Comment | Organism |
---|---|---|
ATP:co(I)rrinoid adenosyltransferase | - |
Salmonella enterica |
CobA | - |
Salmonella enterica |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Salmonella enterica |
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 maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0013 | - |
TWYYGEAQCDWDD68 | 37°C, recombinant wild-type enzyme | Salmonella enterica |