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Literature summary for 1.3.1.124 extracted from
- Studies of human mitochondrial 2,4-dienoyl-CoA reductase (2005), Arch. Biochem. Biophys., 434, 195-200.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| overexpression of His-tagged truncated wild-type and of mutant enzymes in Escherichia coli strain BL21(DE3), expression as soluble protein, except for mutant D117A | Homo sapiens |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D117A | site-directed mutagenesis, mutant is expressed in inclusion bodies and cannot be refolded to a functional enzyme after extraction from inclusion bodies with 8 M urea as denatured protein | Homo sapiens |
| D300A | site-directed mutagenesis, reduced activity and altered kinetics compared to the wild-type enzyme | Homo sapiens |
| E154A | site-directed mutagenesis, reduced activity and altered kinetics compared to the wild-type enzyme | Homo sapiens |
| E227A | site-directed mutagenesis, reduced activity and altered kinetics compared to the wild-type enzyme | Homo sapiens |
| E276A | site-directed mutagenesis, reduced activity and altered kinetics compared to the wild-type enzyme | Homo sapiens |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 3-furan-2-yl-acrylyl-CoA | competitive, substrate analogue | Homo sapiens |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0045 | - |
trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged mutant D300A, pH 6.0, 22°C | Homo sapiens | |
| 0.0065 | - |
2-fluoro-trans-trans-2,4-octadienoyl-CoA | recombinant His-tagged truncated wild-type enzyme, pH 6.0, 22°C | Homo sapiens | |
| 0.008 | - |
NADPH | recombinant His-tagged truncated wild-type enzyme, pH 6.0, 22°C | Homo sapiens | |
| 0.014 | - |
NADPH | recombinant His-tagged mutant E227A, pH 6.0, 22°C | Homo sapiens | |
| 0.014 | - |
trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged truncated wild-type enzyme, pH 6.0, 22°C | Homo sapiens | |
| 0.016 | - |
5-methyl-trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged truncated wild-type enzyme, pH 6.0, 22°C | Homo sapiens | |
| 0.041 | - |
trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged mutant E227A, pH 6.0, 22°C | Homo sapiens | |
| 0.067 | - |
trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged mutant E276A, pH 6.0, 22°C | Homo sapiens | |
| 0.131 | - |
NADPH | recombinant His-tagged mutant E276A, pH 6.0, 22°C | Homo sapiens | |
| 0.154 | - |
trans-trans-2,4-hexadienoyl-CoA | recombinant His-tagged mutant E154A, pH 6.0, 22°C | Homo sapiens | |
| 0.195 | - |
NADPH | recombinant His-tagged mutant D300A, pH 6.0, 22°C | Homo sapiens | |
| 0.215 | - |
NADPH | recombinant His-tagged mutant E154A, pH 6.0, 22°C | Homo sapiens | |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| mitochondrion | - |
Homo sapiens | 5739 | - |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| additional information | Homo sapiens | key enzyme in the beta-oxidation of unsaturated fatty acids, naturally occuring lethal enzyme deficiency | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Homo sapiens | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged truncated wild-type and of mutant enzymes from Escherichia coli strain BL21(DE3) by one-step nickel affinity chromatography, to over 95% purity | Homo sapiens |
Reaction
| Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|
| a (2E)-2-enoyl-CoA + NADP+ = a (2E,4E)-2,4-dienoyl-CoA + NADPH + H+ | reaction mechanism, E276 is the proton donor in catalysis | Homo sapiens |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| mutant D117A is expressed in inclusion bodies and cannot be refolded to a functional enzyme after extraction from inclusion bodies with 8 M urea as denatured protein | Homo sapiens |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| 2-fluoro-trans-trans-2,4-octadienoyl-CoA + NADPH | substrate analogue, recombinant enzyme, reaction mechanism | Homo sapiens | 2-fluoro-trans-3-octenoyl-CoA + NADP+ | - |
? | |
| 5-methyl-trans-trans-2,4-hexadienoyl-CoA + NADPH | substrate analogue, recombinant enzyme | Homo sapiens | 5-methyl-trans-3-hexenoyl-CoA + NADP+ | - |
? | |
| additional information | key enzyme in the beta-oxidation of unsaturated fatty acids, naturally occuring lethal enzyme deficiency | Homo sapiens | ? | - |
? | |
| trans-trans-2,4-hexadienoyl-CoA + NADPH | reaction mechanism | Homo sapiens | trans-3-hexenoyl-CoA + NADP+ | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| 2,4-dienoyl-CoA reductase | - |
Homo sapiens |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 22 | - |
assay at room temperature | Homo sapiens |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 4 | - |
recombinant mutants E227A, E276A, D300A, and D117A | Homo sapiens |
| 5 | - |
recombinant mutant E154A | Homo sapiens |
| 6 | - |
recombinant wild-type enzyme | Homo sapiens |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| NADPH | - |
Homo sapiens | |
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