Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 (RARE) cells | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
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 |
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 |
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 |
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 |
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 |
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 |
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 |
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 |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q92506 | cf EC 1.1.1.62 | - |
Purification (Comment) | Organism |
---|---|
Ni-NTA column chromatography and Superdex 200 gel filtration | Homo sapiens |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.06 | - |
mutant enzyme K173A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
0.44 | - |
mutant enzyme R34A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
0.44 | - |
mutant enzyme R34A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
0.46 | - |
mutant enzyme K152A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
0.5 | - |
mutant enzyme Q126E/R168E/K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
0.65 | - |
mutant enzyme Y169A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
0.85 | - |
mutant enzyme Y169A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
1.2 | - |
mutant enzyme K173A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
1.3 | - |
mutant enzyme K169E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
1.6 | - |
mutant enzyme R168E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
1.7 | - |
mutant enzyme Q126E/K169E, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
2 | - |
mutant enzyme Y169A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
2.4 | - |
mutant enzyme K173A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
2.6 | - |
wild type enzyme, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
3.1 | - |
mutant enzyme Y169A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
3.5 | - |
wild type enzyme, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
4.3 | - |
mutant enzyme Q126E/K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
4.5 | - |
mutant enzyme K173A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
5.4 | - |
mutant enzyme D42A, with 9,10-phenanthrene quinone and NADP+, pH 7.4 and 25°C | Homo sapiens |
5.5 | - |
mutant enzyme K169E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
6.3 | - |
mutant enzyme R168E, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
7.3 | - |
mutant enzyme D42A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
7.9 | - |
mutant enzyme R168E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
8.4 | - |
mutant enzyme Q126E/R168E/K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
14.8 | - |
mutant enzyme Q126E/R168E/K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
15.7 | - |
mutant enzyme K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
17.4 | - |
mutant enzyme Q126E/K169E, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
19.3 | - |
mutant enzyme K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
20 | - |
mutant enzyme D42A, with acetoacetyl-CoA and NADPH, pH 7.4 and 25°C | Homo sapiens |
20.5 | - |
wild type enzyme, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
20.7 | - |
mutant enzyme R34A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
20.8 | - |
wild type enzyme, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
21 | - |
mutant enzyme R34A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
21.7 | - |
mutant enzyme R168E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
22 | - |
mutant enzyme K152A, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
25.5 | - |
mutant enzyme K152A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
31.8 | - |
mutant enzyme Q126E/K169E, with acetoacetyl-CoA and NADH, pH 7.4 and 25°C | Homo sapiens |
74.6 | - |
mutant enzyme D42A, with 9,10-phenanthrene quinone and NAD+, pH 7.4 and 25°C | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
9,10-phenanthrene quinone + NADH + H+ | - |
Homo sapiens | ? | - |
? | |
9,10-phenanthrene quinone + NADPH + H+ | low activity with NADPH | Homo sapiens | ? | - |
? | |
acetoacetyl-CoA + NADH + H+ | - |
Homo sapiens | ? | - |
? | |
acetoacetyl-CoA + NADPH + H+ | low activity with NADPH | Homo sapiens | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
heterotetramer | - |
Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
3-ketoacyl-ACP reductase/3R-hydroxyacyl-CoA dehydrogenase | - |
Homo sapiens |
3-ketoacyl-thioester reductase | - |
Homo sapiens |
KAR | - |
Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADH | - |
Homo sapiens | |
NADPH | low activity with NADPH | Homo sapiens |