Cloned (Comment) | Organism |
---|---|
overexpression of wild-type enzyme and mutant Y73N in enzyme-deficient mutant strain BJ1991 mrf1DELTA, using the catalase 1 promotor | Saccharomyces cerevisiae |
overexpression of wild-type enzyme and mutant Y79N in enzyme-deficient Saccharomyces cerevisiae mutant strain BJ1991 mrf1DELTA, using the catalase 1 promotor | Candida tropicalis |
Crystallization (Comment) | Organism |
---|---|
purified recombinant wild-type enzyme free and in complex with NADPH, mutant enzyme Y79N, hanging drop vapour diffusion method, 22°C, 15-25 mg/ml free enzyme in 50 mM sodium phosphate, pH 7.0, 150 mM NaCl with or without 10 mM NADPH, in a 1:1 ratio with reservoir solution containing 1.9 M ammonium sulfate, and 0.1 M N-(2-acetamido)-2-iminodiacetic acid/NaOH, at pH 6.5 for the free enzyme and pH 7.0 for the cofactor-complexed enzyme, mutant Y79N is crystallized using 15 mg/ml protein and reservoir solution at pH 7.0, labeling by soaking in heavy-atom-solutions, X-ray diffraction structure determination and analysis at 1.7 A, 2.25 A, and 2.6 A, respectively | Candida tropicalis |
Protein Variants | Comment | Organism |
---|---|---|
Y73N | site-directed mutagenesis, mutant shows native fold, but only residual wild-type enzyme activity, no rescue of the enzyme-deficient mutant strain mrf1DELTA | Saccharomyces cerevisiae |
Y79N | site-directed mutagenesis, mutant shows native fold, but only 0.1% of the wild-type enzyme activity, no rescue of the enzyme-deficient mutant strain mrf1DELTA | Candida tropicalis |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
mitochondrion | - |
Candida tropicalis | 5739 | - |
mitochondrion | - |
Saccharomyces cerevisiae | 5739 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | Candida tropicalis | reaction within the fatty acid synthase complex, indispensable for respiratory function in mitochondria | acyl-[acyl-carrier protein] + NADP+ | - |
? | |
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | Saccharomyces cerevisiae | reaction within the fatty acid synthase complex, indispensable for respiratory function in mitochondria | acyl-[acyl-carrier protein] + NADP+ | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Candida tropicalis | - |
enzyme Etr1p | - |
Saccharomyces cerevisiae | P38071 | enzyme Mrf1p | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type enzyme and mutant Y73N from mutant strain BJ1991 mrf1DELTA | Saccharomyces cerevisiae |
recombinant wild-type enzyme and mutant Y79N from Saccharomyces cerevisiae mutant strain BJ1991 mrf1DELTA | Candida tropicalis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.2 | - |
purified recombinant mutant Y79N | Candida tropicalis |
20 | - |
purified recombinant wild-type enzyme | Candida tropicalis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | - |
Candida tropicalis | acyl-[acyl-carrier protein] + NADP+ | - |
? | |
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | - |
Saccharomyces cerevisiae | acyl-[acyl-carrier protein] + NADP+ | - |
? | |
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | reaction within the fatty acid synthase complex, indispensable for respiratory function in mitochondria | Candida tropicalis | acyl-[acyl-carrier protein] + NADP+ | - |
? | |
trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH | reaction within the fatty acid synthase complex, indispensable for respiratory function in mitochondria | Saccharomyces cerevisiae | acyl-[acyl-carrier protein] + NADP+ | - |
? | |
trans-2-hexenoyl-CoA + NADPH | - |
Candida tropicalis | hexanoyl-CoA + NADP+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
2-enoyl thioester reductase | - |
Candida tropicalis |
2-enoyl thioester reductase | - |
Saccharomyces cerevisiae |
enoyl thioester reductase | - |
Candida tropicalis |
enoyl thioester reductase | - |
Saccharomyces cerevisiae |
Etr1p | - |
Candida tropicalis |
More | enzyme belongs to the medium-chain dehydrogenase/reductase superfamily of enzymes | Candida tropicalis |
More | enzyme belongs to the medium-chain dehydrogenase/reductase superfamily of enzymes | Saccharomyces cerevisiae |
Mrf1p | - |
Saccharomyces cerevisiae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | dependent on | Saccharomyces cerevisiae | |
NADPH | dependent on, tyrosine in the binding cleft is critical for enzyme activity, NADPH binding induces conformational changes in the enzyme, overview | Candida tropicalis |