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Literature summary extracted from

  • Venkatesan, R.; Sah-Teli, S.K.; Awoniyi, L.O.; Jiang, G.; Prus, P.; Kastaniotis, A.J.; Hiltunen, J.K.; Wierenga, R.K.; Chen, Z.
    Insights into mitochondrial fatty acid synthesis from the structure of heterotetrameric 3-ketoacyl-ACP reductase/3R-hydroxyacyl-CoA dehydrogenase (2014), Nat. Commun., 5, 4805.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.100 expressed in Escherichia coli BL21 (RARE) cells Homo sapiens

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.100 sitting drop vapor diffusion method, using 15-18% (w/v) PEG3350 and 0.4 M ammonium acetate in sodium acetate buffer at pH 5.0 Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.100 D42A the mutant shows 36% activity with acetoacetyl-CoA and NADH, 20% activity with acetoacetyl-CoA and NADPH, 4% activity with 9,10-phenanthrene and NAD+, and 26% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 K152A the mutant shows 109% activity with acetoacetyl-CoA and NADH, no activity with acetoacetyl-CoA and NADPH, 123% activity with 9,10-phenanthrene and NAD+, and 2.2% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 K169E the mutant shows 95% activity with acetoacetyl-CoA and NADH, 5.5% activity with acetoacetyl-CoA and NADPH, 76% activity with 9,10-phenanthrene and NAD+, and 6% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 K173A the mutant shows 0.3% activity with acetoacetyl-CoA and NADH, 2.4% activity with acetoacetyl-CoA and NADPH, 6% activity with 9,10-phenanthrene and NAD+, and 22% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 Q126E/K169E the mutant shows 156% activity with acetoacetyl-CoA and NADH, 4.3% activity with acetoacetyl-CoA and NADPH, 84% activity with 9,10-phenanthrene and NAD+, and 8% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 Q126E/R168E/K169E the mutant shows 72% activity with acetoacetyl-CoA and NADH, 0.5% activity with acetoacetyl-CoA and NADPH, 40% activity with 9,10-phenanthrene and NAD+, and no activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 R168E the mutant shows 106% activity with acetoacetyl-CoA and NADH, 6.3% activity with acetoacetyl-CoA and NADPH, 38% activity with 9,10-phenanthrene and NAD+, and 8% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 R34A the mutant shows 102% activity with acetoacetyl-CoA and NADH, 100% activity with acetoacetyl-CoA and NADPH, 40.44% activity with 9,10-phenanthrene and NAD+, and 2.1% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens
1.1.1.100 Y169A the mutant shows 3% activity with acetoacetyl-CoA and NADH, 2% activity with acetoacetyl-CoA and NADPH, 4% activity with 9,10-phenanthrene and NAD+, and 15% activity with 9,10-phenanthrene and NADP+, compared to the wild type enzyme, respectively Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.100 Homo sapiens Q92506 cf EC 1.1.1.62
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.100 Ni-NTA column chromatography and Superdex 200 gel filtration Homo sapiens

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.100 0.06
-
 mutant enzyme K173A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.44
-
 mutant enzyme R34A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.44
-
 mutant enzyme R34A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.46
-
 mutant enzyme K152A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.5
-
 mutant enzyme Q126E/R168E/K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.65
-
 mutant enzyme Y169A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 0.85
-
 mutant enzyme Y169A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 1.2
-
 mutant enzyme K173A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 1.3
-
 mutant enzyme K169E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 1.6
-
 mutant enzyme R168E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 1.7
-
 mutant enzyme Q126E/K169E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 2
-
 mutant enzyme Y169A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 2.4
-
 mutant enzyme K173A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 2.6
-
 wild type enzyme, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 3.1
-
 mutant enzyme Y169A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 3.5
-
 wild type enzyme, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 4.3
-
 mutant enzyme Q126E/K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 4.5
-
 mutant enzyme K173A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 5.4
-
 mutant enzyme D42A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 5.5
-
 mutant enzyme K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 6.3
-
 mutant enzyme R168E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 7.3
-
 mutant enzyme D42A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 7.9
-
 mutant enzyme R168E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 8.4
-
 mutant enzyme Q126E/R168E/K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 14.8
-
 mutant enzyme Q126E/R168E/K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 15.7
-
 mutant enzyme K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 17.4
-
 mutant enzyme Q126E/K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 19.3
-
 mutant enzyme K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 20
-
 mutant enzyme D42A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 20.5
-
 wild type enzyme, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 20.7
-
 mutant enzyme R34A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 20.8
-
 wild type enzyme, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 21
-
 mutant enzyme R34A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 21.7
-
 mutant enzyme R168E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 22
-
 mutant enzyme K152A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 25.5
-
 mutant enzyme K152A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens
1.1.1.100 31.8
-
 mutant enzyme Q126E/K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C Homo sapiens
1.1.1.100 74.6
-
 mutant enzyme D42A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.100 9,10-phenanthrene quinone + NADH + H+
-
 Homo sapiens ?
-
 ?
1.1.1.100 9,10-phenanthrene quinone + NADPH + H+ low activity with NADPH Homo sapiens ?
-
 ?
1.1.1.100 acetoacetyl-CoA + NADH + H+
-
 Homo sapiens ?
-
 ?
1.1.1.100 acetoacetyl-CoA + NADPH + H+ low activity with NADPH Homo sapiens ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.100 heterotetramer
-
 Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.100 3-ketoacyl-ACP reductase/3R-hydroxyacyl-CoA dehydrogenase
-
 Homo sapiens
1.1.1.100 3-ketoacyl-thioester reductase
-
 Homo sapiens
1.1.1.100 KAR
-
 Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.100 NADH
-
 Homo sapiens
1.1.1.100 NADPH low activity with NADPH Homo sapiens