Literature summary for 1.3.1.10 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.

Cloned(Commentary)

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 (Commentary)

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

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

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

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

Natural Substrates/ Products (Substrates)

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

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

Purification (Commentary)

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 [micromol/min/mg]

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 and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
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

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

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