BRENDA - Enzyme Database

Role of active site loop in coenzyme binding and flavin reduction in cytochrome P450 reductase

Mothersole, R.G.; Meints, C.E.; Louder, A.; Wolthers, K.R.; Arch. Biochem. Biophys. 606, 111-119 (2016)

Data extracted from this reference:

Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.6.2.4
A635G/R636S
the mutations lead to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
1.6.2.4
D634A
the variant elicits a modest increase in coenzyme binding affinity coupled with a 36fold reduction in cytochrome c turnover and a 17fold decrease in the pre-steady state rate of flavin reduction
Homo sapiens
1.6.2.4
D634N
the variant elicits a modest increase in coenzyme binding affinity coupled with a 10fold reduction in cytochrome c turnover and a 3fold decrease in the pre-steady state rate of flavin reduction
Homo sapiens
1.6.2.4
R636A
the mutation leads to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
1.6.2.4
R636S
the mutation leads to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.6.2.4
2',5'-ADP
-
Homo sapiens
1.6.2.4
NADP+
-
Homo sapiens
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.6.2.4
0.00014
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00019
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0007
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00106
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0016
-
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0018
-
NADPH
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.6.2.4
endoplasmic reticulum
-
Homo sapiens
5783
-
1.6.2.4
membrane
-
Homo sapiens
16020
-
1.6.2.4
nuclear envelope
-
Homo sapiens
5635
-
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.6.2.4
Homo sapiens
P16435
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.6.2.4
Ni-NTA column chromatography
Homo sapiens
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.2.4
NADPH + H+ + oxidized cytochrome c
-
741783
Homo sapiens
NADP+ + reduced cytochrome c
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.6.2.4
?
x * 78000, calculated from amino acid sequence
Homo sapiens
Turnover Number [1/s]
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.6.2.4
0.56
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
2
8
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
2.21
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
20
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
23.4
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
30.5
-
NADPH
mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
38.6
-
NADPH
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.6.2.4
FAD
-
Homo sapiens
1.6.2.4
FMN
-
Homo sapiens
Ki Value [mM]
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.6.2.4
0.00037
-
2',5'-ADP
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00041
-
2',5'-ADP
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00043
-
2',5'-ADP
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0006
-
2',5'-ADP
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00066
-
NADP+
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00072
-
NADP+
mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00074
-
NADP+
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.001
-
NADP+
mutant enzyme A635G, at pH 7.5 and 25°C; wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00131
-
2',5'-ADP
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00174
-
NADP+
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0027
-
NADP+
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0045
-
2',5'-ADP
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.6.2.4
FAD
-
Homo sapiens
1.6.2.4
FMN
-
Homo sapiens
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.6.2.4
A635G/R636S
the mutations lead to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
1.6.2.4
D634A
the variant elicits a modest increase in coenzyme binding affinity coupled with a 36fold reduction in cytochrome c turnover and a 17fold decrease in the pre-steady state rate of flavin reduction
Homo sapiens
1.6.2.4
D634N
the variant elicits a modest increase in coenzyme binding affinity coupled with a 10fold reduction in cytochrome c turnover and a 3fold decrease in the pre-steady state rate of flavin reduction
Homo sapiens
1.6.2.4
R636A
the mutation leads to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
1.6.2.4
R636S
the mutation leads to a modest increase in cytochrome c reduction, which is linked to weaker coenzyme binding and faster interflavin electron transfer
Homo sapiens
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.6.2.4
2',5'-ADP
-
Homo sapiens
1.6.2.4
NADP+
-
Homo sapiens
Ki Value [mM] (protein specific)
EC Number
Ki Value [mM]
Ki Value maximum [mM]
Inhibitor
Commentary
Organism
Structure
1.6.2.4
0.00037
-
2',5'-ADP
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00041
-
2',5'-ADP
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00043
-
2',5'-ADP
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0006
-
2',5'-ADP
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00066
-
NADP+
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00072
-
NADP+
mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00074
-
NADP+
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.001
-
NADP+
mutant enzyme A635G, at pH 7.5 and 25°C; wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00131
-
2',5'-ADP
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00174
-
NADP+
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0027
-
NADP+
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0045
-
2',5'-ADP
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.6.2.4
0.00014
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00019
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0007
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.00106
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0016
-
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
0.0018
-
NADPH
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.6.2.4
endoplasmic reticulum
-
Homo sapiens
5783
-
1.6.2.4
membrane
-
Homo sapiens
16020
-
1.6.2.4
nuclear envelope
-
Homo sapiens
5635
-
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.6.2.4
Ni-NTA column chromatography
Homo sapiens
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.2.4
NADPH + H+ + oxidized cytochrome c
-
741783
Homo sapiens
NADP+ + reduced cytochrome c
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.6.2.4
?
x * 78000, calculated from amino acid sequence
Homo sapiens
Turnover Number [1/s] (protein specific)
EC Number
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
1.6.2.4
0.56
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
2
8
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
2.21
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
20
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
23.4
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
30.5
-
NADPH
mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
38.6
-
NADPH
mutant enzyme A635G/R636S, at pH 7.5 and 25°C
Homo sapiens
KCat/KM [mM/s]
EC Number
kcat/KM Value [1/mMs-1]
kcat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.6.2.4
2900
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
15800
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
17500
-
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
22100
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme A635G/R636S, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
28200
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens
KCat/KM [mM/s] (protein specific)
EC Number
KCat/KM Value [1/mMs-1]
KCat/KM Value Maximum [1/mMs-1]
Substrate
Commentary
Organism
Structure
1.6.2.4
2900
-
NADPH
mutant enzyme D634A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
15800
-
NADPH
mutant enzyme D634N, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
17500
-
NADPH
mutant enzyme R636A, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
22100
-
NADPH
mutant enzyme A635G, at pH 7.5 and 25°C; mutant enzyme A635G/R636S, at pH 7.5 and 25°C; mutant enzyme R636S, at pH 7.5 and 25°C
Homo sapiens
1.6.2.4
28200
-
NADPH
wild type enzyme, at pH 7.5 and 25°C
Homo sapiens