Cloned (Comment) | Organism |
---|---|
overexpression of the His-tagged enzyme in Escherichia coli as soluble protein, expression of enzyme for selenomethionine-labeling in Escherichia coli strain B834(DE3) | Mus musculus |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged wild-type and mutants M335V and C323M enzymes free or in complex with octanoylcarnitine, sitting drop vapour diffusion method, 4°C, reservoir solution contains 0.1 M HEPES, pH 7.4, 62% v/v 2-methyl-2,4-pentanediol, larger crystals by micro- and macroseeding, cross-seeding of recombinant purified selenomethionine-labeled enzyme, X-ray diffraction structure determination and analysis at 2.0-2.2 A resolution | Mus musculus |
Protein Variants | Comment | Organism |
---|---|---|
C323M | site-directed mutagenesis, crystal structure comparison with the wild-type enzyme, altered acyl-CoA substrate specificity compared to the wild-type enzyme, highly increased specificity for octanoyl-CoA | Mus musculus |
G553M | site-directed mutagenesis, highly reduced activity with altered acyl-CoA substrate specificity compared to the wild-type enzyme | Mus musculus |
M335A | site-directed mutagenesis, reduced activity with altered acyl-CoA substrate specificity compared to the wild-type enzyme | Mus musculus |
M335V | site-directed mutagenesis, crystal structure comparison with the wild-type enzyme, highly reduced activity with altered acyl-CoA substrate specificity compared to the wild-type enzyme | Mus musculus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
octanoyl-CoA + L-carnitine | Mus musculus | - |
CoA + L-octanoylcarnitine | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Mus musculus | Q9DC50 | - |
- |
Purification (Comment) | Organism |
---|---|
soluble recombinant His-tagged wild-type and selenomethionine-labeled enzyme from Escherichia coli by nickel affinity chromatography | Mus musculus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
octanoyl-CoA + L-carnitine = CoA + L-octanoylcarnitine | acyl-binding hydrophobic pocket structure involving residues G553, C323, and Met335, the acyl-CoA substrate chain length specificity is determined by G553, Met335 is important for catalysis | Mus musculus |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
substrate specificity of wild-type and mutant enzymes, overview | Mus musculus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
acetyl-CoA + L-carnitine | low activity | Mus musculus | CoA + L-acetylcarnitine | - |
? | |
octanoyl-CoA + L-carnitine | - |
Mus musculus | CoA + L-octanoylcarnitine | - |
r | |
octanoyl-CoA + L-carnitine | preferred substrate | Mus musculus | CoA + L-octanoylcarnitine | - |
r | |
tetradecanoyl-CoA + L-carnitine | low activity | Mus musculus | CoA + L-tetradecanoylcarnitine | - |
? |
Synonyms | Comment | Organism |
---|---|---|
carnitine octanoyltransferase | - |
Mus musculus |
CrOT | - |
Mus musculus |
More | the enzyme belongs to the carnitine acyltransferase family | Mus musculus |