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Literature summary extracted from

  • Airenne, T.T.; Torkko, J.M.; Van den plas, S.; Sormunen, R.T.; Kastaniotis, A.J.; Wierenga, R.K.; Kalervo Hiltunen, J.
    Structure-function analysis of enoyl thioester reductase involved in mitochondrial maintenance (2003), J. Mol. Biol., 327, 47-59.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.3.1.10 overexpression of wild-type enzyme and mutant Y73N in enzyme-deficient mutant strain BJ1991 mrf1DELTA, using the catalase 1 promotor Saccharomyces cerevisiae
1.3.1.10 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 (Commentary)

EC Number Crystallization (Comment) Organism
1.3.1.10 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

EC Number Protein Variants Comment Organism
1.3.1.10 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
1.3.1.10 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

EC Number Localization Comment Organism GeneOntology No. Textmining
1.3.1.10 mitochondrion
-
 Candida tropicalis 5739
-
1.3.1.10 mitochondrion
-
 Saccharomyces cerevisiae 5739
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.3.1.10 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+
-
 ?
1.3.1.10 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

EC Number Organism UniProt Comment Textmining
1.3.1.10 Candida tropicalis
-
 enzyme Etr1p
-
1.3.1.10 Saccharomyces cerevisiae P38071 enzyme Mrf1p
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.3.1.10 recombinant wild-type enzyme and mutant Y73N from mutant strain BJ1991 mrf1DELTA Saccharomyces cerevisiae
1.3.1.10 recombinant wild-type enzyme and mutant Y79N from Saccharomyces cerevisiae mutant strain BJ1991 mrf1DELTA Candida tropicalis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.3.1.10 0.2
-
 purified recombinant mutant Y79N Candida tropicalis
1.3.1.10 20
-
 purified recombinant wild-type enzyme Candida tropicalis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.3.1.10 trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH
-
 Candida tropicalis acyl-[acyl-carrier protein] + NADP+
-
 ?
1.3.1.10 trans-2,3-dehydroacyl-[acyl-carrier protein] + NADPH
-
 Saccharomyces cerevisiae acyl-[acyl-carrier protein] + NADP+
-
 ?
1.3.1.10 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+
-
 ?
1.3.1.10 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+
-
 ?
1.3.1.10 trans-2-hexenoyl-CoA + NADPH
-
 Candida tropicalis hexanoyl-CoA + NADP+
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.3.1.10 2-enoyl thioester reductase
-
 Candida tropicalis
1.3.1.10 2-enoyl thioester reductase
-
 Saccharomyces cerevisiae
1.3.1.10 enoyl thioester reductase
-
 Candida tropicalis
1.3.1.10 enoyl thioester reductase
-
 Saccharomyces cerevisiae
1.3.1.10 Etr1p
-
 Candida tropicalis
1.3.1.10 More enzyme belongs to the medium-chain dehydrogenase/reductase superfamily of enzymes Candida tropicalis
1.3.1.10 More enzyme belongs to the medium-chain dehydrogenase/reductase superfamily of enzymes Saccharomyces cerevisiae
1.3.1.10 Mrf1p
-
 Saccharomyces cerevisiae

Cofactor

EC Number Cofactor Comment Organism Structure
1.3.1.10 NADPH dependent on Saccharomyces cerevisiae
1.3.1.10 NADPH dependent on, tyrosine in the binding cleft is critical for enzyme activity, NADPH binding induces conformational changes in the enzyme, overview Candida tropicalis