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+ NADPH + H+
? + NADP+
-
-
-
?
1,4-dioxo-2-(trifluoromethyl)-1lambda5,4lambda5-quinoxaline + NADPH + H+
? + NADP+
-
-
-
?
1,4-dioxo-7-(trifluoromethoxy)-1lambda5,2,4lambda5-benzotriazin-3-amine + NADPH + H+
? + NADP+
-
-
-
?
1-oxo-1lambda5,2,4-benzotriazine + NADPH + H+
? + NADP+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
2 ferricytochrome P450 + NADPH
2 ferrocytochrome P450 + NADP+ + H+
-
-
-
?
2 oxidized nitroblue tetrazolium + NADPH
2 reduced nitroblue tetrazolium + NADP+ + H+
-
-
-
?
3-amino-1,4-dioxo-1lambda5,4lambda5-quinoxaline-2-carbonitrile + NADPH + H+
? + NADP+
-
-
-
?
methyl (1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)carbamate + NADPH + H+
? + NADP+
-
-
-
?
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)-1,1,1-trifluoromethanesulfonamide + NADPH + H+
? + NADP+
-
-
-
?
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)acetamide + NADPH + H+
? + NADP+
-
-
-
?
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)methanesulfonamide + NADPH + H+
? + NADP+
-
-
-
?
N-(1-oxo-1lambda5,2,4-benzotriazin-3-yl)acetamide + NADPH + H+
? + NADP+
-
-
-
?
NADPH + H+ + 2 oxidized cytochrome c
NADP+ + 2 reduced cytochrome c
-
-
-
?
NADPH + H+ + 2 oxidized cytochrome P450 2B4
NADP+ + 2 reduced cytochrome P450 2B4
-
-
-
?
NADPH + H+ + cytochrome c
NADP+ + reduced cytochrome c
-
-
-
?
NADPH + H+ + ferricytochrome c
NADP+ + ferrocytochrome c
-
-
-
?
tirapazamine + NADPH + H+
1,2,4-benzotriazin-3-amine + 1-oxo-1lambda5,2,4-benzotriazin-3-amine + NADP+
-
-
-
?
2 ferricyanide + NADPH
2 ferrocyanide + NADP+ + H+
-
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
2 ferricytochrome P450 + NADPH
2 ferrocytochrome P450 + NADP+ + H+
NADPH + 5-cyano-2,3-di-p-tolyltetrazolium chloride
NADP+ + (5-cyano-2,3-di-p-tolyltetrazolium chloride) formazan
-
-
formazan is fluorescent
-
?
NADPH + H+ + 2,4,6-trinitrotoluene
NADP+ + nitrite + ?
-
-
-
-
?
NADPH + H+ + 4-nitroacetophenone
?
-
-
-
-
r
NADPH + H+ + 4-nitrobenzaldehyde
?
-
-
-
-
r
NADPH + H+ + 4-nitroso-2,6-dinitrotoluene
NADP+ + 4-hydroxyamino-2,6-dinitrotoluene
-
-
-
-
?
NADPH + H+ + benzalacetone
?
-
-
-
-
r
NADPH + H+ + cytochrome c
NADP+ + reduced cytochrome c
-
-
-
-
?
NADPH + H+ + oxidized 2,6-dichlorophenolindophenol
NADP+ + reduced 2,6-dichlorophenolindophenol
-
-
-
-
?
NADPH + H+ + oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
NADP+ + reduced 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
-
-
-
?
NADPH + H+ + oxidized cytochrome c
NADP+ + reduced cytochrome c
-
-
-
-
?
NADPH + H+ + oxidized menadione
NADP+ + reduced menadione
-
-
-
-
?
NADPH + hexadecanal
NADP+ + hexadecanol
-
brain, hexadecanal replaceable by p-nitroacetophenone, or p-pyridinecarboxaldehyde, benzalacetone or p-nitrobenzaldehyde
-
?
NADPH + octanal
NADP+ + octanol
-
-
-
?
additional information
?
-
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
-
-
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: ferricyanide
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: ferricyanide
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: ferricyanide
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: ferricyanide
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: ferricyanide
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome b5
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: neotetrazolium chloride
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: neotetrazolium chloride
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: benzoquinone
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: vitamin K3
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: formylated cytochrome c
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: cytochrome P450
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: 2,6-dichlorophenolindophenol
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: 2,6-dichlorophenolindophenol
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: 2,6-dichlorophenolindophenol
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
additional electron acceptor: 2,6-dichlorophenolindophenol
-
?
2 ferricytochrome c + NADPH
2 ferrocytochrome c + NADP+ + H+
-
NADPH-cytochrome P450 reductase is a unique universal donor of electrons to practically all known microsomal cytochrome P450s and one of the most important components of the monooxygenase system
-
-
?
2 ferricytochrome P450 + NADPH
2 ferrocytochrome P450 + NADP+ + H+
-
-
-
-
?
2 ferricytochrome P450 + NADPH
2 ferrocytochrome P450 + NADP+ + H+
-
CPR shuttles electrons from NADPH through the FAD and FMN-coenzymes into the iron of the prosthetic heme-group of the cytochrome P450
-
-
?
additional information
?
-
the presence of NADPH-cytochrome P450 reductase prevents heme oxygenase-1 from acetylation of lysine residues, Lys149 and Lys153, located in the F-helix. The heme degradation activity of the fully acetylated heme oxygenase-1 in the NADPH/NADPH-cytochrome P450 reductase-supported system is significantly reduced, whereas almost no inactivation is detected in heme oxygenase-1 in the presence of NADPH-cytochrome P450 reductase, which prevents acetylation of Lys149 and Lys153
-
-
?
additional information
?
-
-
hydroxylation of benzo-a-pyrene, testosterone and progesterone
-
-
?
additional information
?
-
-
denitration of glyceryl trinitrate
-
-
?
additional information
?
-
-
O-dealkylation of pentoxyresorufin
-
-
?
additional information
?
-
-
component of the electron transport chain required for activity of the 17,20-lyase in testis microsomes, removal of 2-carbon side chain from 17-position of 21-carbon steroids
-
-
?
additional information
?
-
-
involved in the reduction of testosterone
-
-
?
additional information
?
-
-
as part of the microsomal ethanol-oxidizing system composed of NADPH-cytochrome c reductase, cytochrome P-450, phospholipids
-
-
?
additional information
?
-
-
aniline hydroxylase
-
-
?
additional information
?
-
-
N-demethylation of aminopyrine
-
-
?
additional information
?
-
-
N-demethylation of benzphetamine
-
-
?
additional information
?
-
-
N-demethylation of benzphetamine
-
-
?
additional information
?
-
-
detoxification of drugs, inactivation of procarcinogens
-
-
?
additional information
?
-
-
monooxygenase system composed of cytochrome P-450, NADPH-cytochrome c reductase, phospholipids
-
-
?
additional information
?
-
-
microsomal heme oxygenase system composed of heme oxygenase and NADPH-cytochrome c reductase catalyzes the oxidative degradation of heme to biliverdin, essential role in the physiological heme catabolism
-
-
?
additional information
?
-
-
the enzyme also catalyzes the oxidative deformylation of a model xenobiotic aldehyde, 2-phenylpropionaldehyde, to the n-1 alcohol, 1-phenylethanol in the absence of cytochrome P450
-
-
?
additional information
?
-
-
the C-terminal 23 amino acids of heme oxygenase-1 play an important role in the interaction between heme oxygenase-1 and NADPH cytochrome P450 reductase, enhancing the conversion of hemin to biliverdin
-
-
?
additional information
?
-
-
cytochrome c is the most popular electron acceptor, several dyes and electron acceptors are also in use: dichlorophenol indophenol, ferricyanide and tetrazoliums
-
-
?
additional information
?
-
-
NADPH-cytochrome P450 reductase interacts with heme oxygenase-1
-
-
?
additional information
?
-
-
2,4,6-trinitrophenol, 2,4-dinitrophenol, 2,6-dinitrotoluene, 1,2-dinitrobenzene, 1,4-dinitrobenzene, 2,4-dinitrobenzoic acid, 3,5-dinitrobenzoic acid, 2-nitrotoluene, 2-nitrobenzonitrile, 4-nitrotoluene and 4-nitrobenzaldehyde are not subject to denitration by the enzyme
-
-
?
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0.000001 - 0.0251
cytochrome c
1.4
4-Nitroacetophenone
-
-
0.31
4-nitrobenzaldehyde
-
-
0.05
5-cyano-2,3-ditolyl tetrazolium chloride
-
-
0.013 - 0.0211
cytochrome c
7.2
ethanol
-
microsomal ethanol oxidizing system
0.002 - 0.026
ferricytochrome c
0.0018 - 0.011
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
additional information
additional information
-
0.000001
cytochrome c
wild type, cosubstrate FAD, Km below
0.00012
cytochrome c
T491V mutant, cosubstrate FAD
0.0008
cytochrome c
G488L mutant, cosubstrate FAD
0.0015
cytochrome c
W677X mutant, cosubstrate NADPH
0.0028
cytochrome c
W677Y mutant, cosubstrate NADPH
0.0041
cytochrome c
R454E mutant, cosubstrate NADPH
0.0056
cytochrome c
S678X mutant, cosubstrate NADPH
0.0058
cytochrome c
Y456S mutant, cosubstrate NADPH
0.0059
cytochrome c
T491V mutant, cosubstrate NADPH
0.0062
cytochrome c
wild type, cosubstrate NADPH
0.0063
cytochrome c
C472T mutant, cosubstrate NADPH
0.0077
cytochrome c
G488L mutant, cosubstrate NADPH
0.0078
cytochrome c
Y456S mutant, cosubstrate FAD
0.0135
cytochrome c
deletion mutant T236/G237
0.015
cytochrome c
deletion mutant T236/G237/E238/E239
0.0251
cytochrome c
R454E mutant, cosubstrate FAD
0.000001
NADPH
wild type, cosubstrate FAD, Km below
0.00012
NADPH
T491V mutant, cosubstrate FAD
0.0008
NADPH
G488L mutant, cosubstrate FAD
0.0027
NADPH
deletion mutant T236/G237/E238/E239
0.003
NADPH
deletion mutant T236/G237
0.0078
NADPH
Y456S mutant, cosubstrate FAD
0.0085
NADPH
W677X mutant, cosubstrate cytochrome c
0.0134
NADPH
G488L mutant, cosubstrate cytochrome c
0.0143
NADPH
W677Y mutant, cosubstrate cytochrome c
0.0163
NADPH
wild type, cosubstrate cytochrome c
0.0166
NADPH
R454E mutant, cosubstrate cytochrome c
0.0199
NADPH
S678X mutant, cosubstrate cytochrome c
0.0201
NADPH
T491V mutant, cosubstrate cytochrome c
0.0217
NADPH
C472T mutant, cosubstrate cytochrome c
0.0251
NADPH
R454E mutant, cosubstrate FAD
0.0548
NADPH
Y456S mutant, cosubstrate cytochrome c
0.013
cytochrome c
-
-
0.0131
cytochrome c
-
Y140F/178F double mutant
0.014
cytochrome c
-
Y178F mutant
0.0178
cytochrome c
-
native enzyme, liver
0.0185
cytochrome c
-
Y178D mutant
0.0188
cytochrome c
-
Y140F mutant
0.0191
cytochrome c
-
Y140D mutant
0.0211
cytochrome c
-
wild type
0.002
ferricytochrome c
-
wild type enzyme, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0024
ferricytochrome c
-
mutant enzyme E115A/E116A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0025
ferricytochrome c
-
mutant enzyme D113A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0036
ferricytochrome c
-
mutant enzyme E115A/E116A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.004
ferricytochrome c
-
wild type enzyme, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0052
ferricytochrome c
-
mutant enzyme D113A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.006
ferricytochrome c
-
mutant enzyme E115A/E116A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0085
ferricytochrome c
-
wild type enzyme, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.011
ferricytochrome c
-
mutant enzyme D113A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.013
ferricytochrome c
-
mutant enzyme E115A/E116A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.016
ferricytochrome c
-
mutant enzyme E115A/E116A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.02
ferricytochrome c
-
wild type enzyme, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.023
ferricytochrome c
-
wild type enzyme, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.025
ferricytochrome c
-
mutant enzyme D113A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.026
ferricytochrome c
-
mutant enzyme D113A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.001 - 0.003
NADPH
-
-
0.0064
NADPH
-
wild type, cosubstrate cytochrome c
0.0066
NADPH
-
native enzyme, liver, cosubstrate cytochrome c
0.0066
NADPH
-
Y140D mutant, cosubstrate cytochrome c
0.0071
NADPH
-
Y178F mutant, cosubstrate cytochrome c
0.0078
NADPH
-
Y140F mutant, cosubstrate cytochrome c
0.0095
NADPH
-
Y178D mutant, cosubstrate cytochrome c
0.0114
NADPH
-
Y140F/178F double mutant, cosubstrate cytochrome c
0.0018
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0021
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0022
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0026
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0026
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0031
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0038
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0039
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0041
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0062
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0063
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0065
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.0087
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.01
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
0.011
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
additional information
additional information
steady-state kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
kinetics by stopped-flow spectroscopy
-
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42
5-cyano-2,3-ditolyl tetrazolium chloride
-
-
14.67 - 65
ferricytochrome c
5.83 - 56.67
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
additional information
additional information
-
20.8 - 22.5
cytochrome c
-
-
21.3
cytochrome c
-
aortic microsomal enzyme
21.9
cytochrome c
-
purified enzyme
14.67
ferricytochrome c
-
wild type enzyme, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
16.67
ferricytochrome c
-
wild type enzyme, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
20
ferricytochrome c
-
wild type enzyme, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
23.3
ferricytochrome c
-
mutant enzyme D113A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
25
ferricytochrome c
-
mutant enzyme D113A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
25
ferricytochrome c
-
mutant enzyme E115A/E116A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
25
ferricytochrome c
-
mutant enzyme E115A/E116A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
33.33
ferricytochrome c
-
wild type enzyme, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
35
ferricytochrome c
-
wild type enzyme, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
36.7
ferricytochrome c
-
mutant enzyme D113A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
36.7
ferricytochrome c
-
mutant enzyme E115A/E116A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
55
ferricytochrome c
-
mutant enzyme D113A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
58.3
ferricytochrome c
-
mutant enzyme E115A/E116A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
58.3
ferricytochrome c
-
mutant enzyme E115A/E116A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
65
ferricytochrome c
-
mutant enzyme D113A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
5.83
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
8.5
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
10.5
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
11.67
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
13
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
13.83
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
18.3
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
18.3
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
20
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
25
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
28.3
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
30
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
38.3
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
40
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
56.67
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
additional information
additional information
-
-
-
additional information
additional information
-
-
-
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27
1,4-dioxo-2-(trifluoromethyl)-1lambda5,4lambda5-quinoxaline
pH 7.4, 25°C
-
46
1,4-dioxo-7-(trifluoromethoxy)-1lambda5,2,4lambda5-benzotriazin-3-amine
pH 7.4, 25°C
-
17
1-oxo-1lambda5,2,4-benzotriazine
pH 7.4, 25°C
-
4.7
3-amino-1,4-dioxo-1lambda5,4lambda5-quinoxaline-2-carbonitrile
pH 7.4, 25°C
-
80
methyl (1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)carbamate
pH 7.4, 25°C
-
25
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)-1,1,1-trifluoromethanesulfonamide
pH 7.4, 25°C
-
70
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)acetamide
pH 7.4, 25°C
-
2.7
N-(1,4-dioxo-1lambda5,2,4lambda5-benzotriazin-3-yl)methanesulfonamide
pH 7.4, 25°C
-
8.7
N-(1-oxo-1lambda5,2,4-benzotriazin-3-yl)acetamide
pH 7.4, 25°C
-
1500 - 10500
ferricytochrome c
2700 - 7200
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
1500
ferricytochrome c
-
wild type enzyme, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
1700
ferricytochrome c
-
wild type enzyme, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2200
ferricytochrome c
-
mutant enzyme D113A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2300
ferricytochrome c
-
wild type enzyme, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2500
ferricytochrome c
-
mutant enzyme D113A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
3300
ferricytochrome c
-
mutant enzyme D113A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
3700
ferricytochrome c
-
mutant enzyme E115A/E116A, using 310 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
3700
ferricytochrome c
-
wild type enzyme, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
4500
ferricytochrome c
-
mutant enzyme D113A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
4500
ferricytochrome c
-
mutant enzyme E115A/E116A, using 210 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
6000
ferricytochrome c
-
mutant enzyme E115A/E116A, using 110 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
7000
ferricytochrome c
-
mutant enzyme E115A/E116A, using 60 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
8300
ferricytochrome c
-
wild type enzyme, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
10000
ferricytochrome c
-
mutant enzyme D113A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
10500
ferricytochrome c
-
mutant enzyme E115A/E116A, using 30 mM ferricytochrome c as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2700
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2700
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
2800
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
3000
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
3000
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
wild type enzyme, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
4700
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
4800
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
4800
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
5000
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
5000
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 210 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
5300
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme D113A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
5800
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 110 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
6500
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 60 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
6800
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 30 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
7200
oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
-
mutant enzyme E115A/E116A, using 10 mM oxidized 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide as substrate, in 20 mM HEPES buffer (pH 7.5), at 25°C
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0.17
R454E mutant, electron acceptor: cytochrome c
0.23
Y456S mutant, electron acceptor: cytochrome c
1.35
G488L mutant, electron acceptor: cytochrome c
1.7
W677X mutant, electron acceptor: cytochrome c
32.7
T491V mutant, electron acceptor: cytochrome c
36
W677Y mutant, electron acceptor: cytochrome c
57.4
wild type, electron acceptor: cytochrome c
62.5
S678X mutant, electron acceptor: cytochrome c
65.4
C472T mutant, electron acceptor: cytochrome c
0.0104
-
aortic microsomal preparation
0.021
-
riboflavin-deficient animals, feeding period 7 weeks, electron acceptor: cytochrome c
0.025
-
electron donor: NADH, electron acceptor: dichlorophenolindophenol
0.03
-
electron donor: NADPH, electron acceptor: cytochrome b5
0.034
-
beta-naphthoflavone-treated animals
0.036
-
control microsomes
0.055
-
electron donor: NADH, electron acceptor: cytochrome c
0.06572
-
purified microsomal ethanol-oxidizing system fraction
0.066
-
riboflavin-deficient animals, feeding period 6 weeks, electron acceptor: cytochrome c
0.075
-
riboflavin deficient animals, feeding period 7 weeks, electron acceptor: ferricyanide
0.097
-
riboflavin-deficient-animals, feeding period 6 weeks, electron acceptor: ferricyanide
0.157
-
control animals, feeding period 6 weeks, electron acceptor: ferricyanide
0.165
-
control animals, feeding period 6 weeks, electron acceptor: cytochrome c
0.189
-
electron acceptor: formylated cytochrome c
0.211
-
brain, electron acceptor: hexadecanal
0.28
-
Y140D/178D double mutant, electron acceptor: cytochrome c
0.44
-
Y178D mutant, electron acceptor: cytochrome c
0.58
-
peroxidation of microsomes for 18 min
0.676
-
purified enzyme, liver
0.72
-
control microsomes
0.928
-
electron acceptor: cytochrome c
11
-
Y140D mutant, electron acceptor: cytochrome c
113
-
Y178F mutant, electron acceptor: ferricyanide
16.5
-
cDNA-expressed liver enzyme
17.9
-
electron donor: NADPH, electron acceptor: dichlorophenolindophenol
41.2
-
electron donor: NADPH, electron acceptor: cytochrome c
46
-
Y140F/178F double mutant, electron acceptor: cytochrome c
47.4
-
Y178F mutant, electron acceptor: cytochrome c
48.1
-
Y178D mutant, electron acceptor: ferricyanide
5.9
-
brain, electron acceptor: p-nitrobenzaldehyde
51
-
Y140D/178D double mutant, electron acceptor: ferricyanide
51.5
-
bacterially expressed reductase protein, electron acceptor: cytochrome c
52.9
-
electron acceptor: ferricyanide
53.3
-
electron acceptor: cytochrome c
55.2
-
Y140F mutant, electron acceptor: cytochrome c
60
-
purified enzyme, microsomes
62.5
-
purification by affinity chromatography on agarose-hexane-adenosine 2',5'-diphosphate
63.8
-
purification by n-octylamino-Sepharose 4B and 2',5'-ADP column chromatography
94.2
-
Y140F/178F double mutant, electron acceptor: ferricyanide
94.8
-
Y140F mutant, electron acceptor: ferricyanide
102
-
bacterially expressed reductase protein, electron acceptor: ferricyanide
102
-
Y140D mutant, electron acceptor: ferricyanide
40.5
-
-
40.5
-
electron donor: NADPH, electron acceptor: ferricyanide
additional information
The substitution mutants (E238A/E239A and T236A/G237A/E238A/E239A) exhibit wild-type activity with cytochrome c. The 2- and 4-alanine addition mutants exhibit about 50% increase in activity compared with the wild-type enzyme. The two and four amino acid deletion mutants reduce cytochrome c 6.4- and 214-fold slower than wild type, respectively.
additional information
-
-
additional information
-
-
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C136A
site-directed mutagenesis
C228A
site-directed mutagenesis
C363T
site-directed mutagenesis
C445L
site-directed mutagenesis
C566A
site-directed mutagenesis, the mutant shows full catalytic activity and a 2.5fold increased Km for NADPH compared to the wild-type enzyme
E238A/E239A
hinge region connecting the FMN domain to the rest of the protein
G237/+AA/E238
hinge region connecting the FMN domain to the rest of the protein
G237/+AAAA/238
hinge region connecting the FMN domain to the rest of the protein
G488L
substitution decreases FAD binding by approximately 80% but does not affect FMN incorporation, 42fold decrease in catalytic activity compared to wild type, substitution does not affect either Km for NADPH or Km for cytochrome c, addition of FAD to the mutant results in partial restoration of catalytic activity
R454E
substitution decreases both FAD binding and FMN incorporation, suggesting interaction between the two flavin domains and/or the interconnecting region, FAD content ranged from undetectable to approximately 0.1 mol of FAD/mol of enzyme, 338fold decrease in catalytic activity compared to wild type, substitution does not affect either Km for NADPH or Km for cytochrome c, addition of FAD to the mutant resulted in partial restoration of catalytic activity
S457A/C630A/D675N
site-directed mutagenesis, catalytically inactive mutant possessing a structure almost identical to that of the wild-type
S678X
substitution does not affect FAD or FMN incorporation, substitution has no effect on the catalytic activity or kinetic properties
T236A/G237A/E238A/E239A
hinge region connecting the FMN domain to the rest of the protein
T491V
substitution decreases FAD binding by approximately 50% but does not affect FMN incorporation, 2fold decrease in catalytic activity compared to wild type, substitution does not affect either Km for NADPH or Km for cytochrome c, addition of FAD to the mutant results in full restoration of catalytic activity
W677X
substitution does not affect FAD or FMN incorporation, 34fold decrease in catalytic activity compared to wild type, substitution does not alter significantly Km for cytochrome c but decreases Km for NADPH
W677Y
substitution does not affect FAD or FMN incorporation, 2fold decrease in catalytic activity compared to wild type, substitution does not alter significantly Km for cytochrome c but decreases Km for NADPH
Y456S
substitution decreases FAD binding but did not affect FMN incorporation, 250fold decrease in catalytic activity compared to wild type, substitution increases Km for cytochrome c, addition of FAD to the mutant results in full restoration of catalytic activity
D113A
-
the mutation increases kcat approximately 2fold, but does not affect Km at the lowest ionic strength (10 mM), the mutant displays no change in catalytic efficiency compared to the wild type enzyme
E115A/E116A
-
the mutations increase kcat approximately 2fold, but does not affect Km at the lowest ionic strength (10 mM), the mutant displays a slight decrease in catalytic efficiency at higher ionic strengths due to a larger increase in Km than observed for kcat
K56Q
-
the full-length mutant enzyme is stable to spontaneopus proteolysis but possesses spectral and catzalytic properties of the wild-type flavoprotein
Y140D
-
substitution does not eliminate FMN binding but reduces cytochrome c reductase activity, Km value for cytochrome c or NADPH similar to wild type
Y140D/178D
-
substitution abolishes FMN binding and cytochrome c reductase activity
Y140F
-
substitution has no effect on FMN content or catalytic activity, Km value for cytochrome c or NADPH similar to wild type
Y140F/178F
-
substitution has no effect on FMN content or catalytic activity, slightly decreases Km for cytochrome c, NADPH Km value slightly higher than wild type
Y178D
-
substitution abolishes FMN binding and cytochrome c reductase activity, Km value for cytochrome c similar to wild type, NADPH Km value slightly higher than wild type
Y178F
-
substitution has no effect on FMN content or catalytic activity, slightly decreases Km for cytochrome c, NADPH Km value similar to wild type
C472T
site-directed mutagenesis
C472T
substitution does not affect FAD or FMN incorporation, substitution has no effect on activity, Km for NADPH or Km for cytochrome c
additional information
deletion mutants in the hinge region (connecting the FMN domain to the rest of the protein): deletion of T236 and G237 or deletion of T236, G237, E238 and E239, also in combination with deletion of the first 56 residues of the N-terminus (resulting in a soluble protein)
additional information
generation of a truncated -56 mutant form W677X of the rat 147CC514, with Trp677 and Ser678 truncated, the mutant exhibits decreased NADP+ binding and alterations in the conformation of the NADP+-binding site
additional information
-
FMN-depleted enzyme is prepared by dialyzing the mutant enzyme Y140A/Y178A, against 2 M KBr. FMN-depleted enzyme can support the conversion of verdoheme to the ferric biliverdin-iron chelate, indicating that electrons required for verdoheme oxidation can be transferred through a pathway not involving FMN
additional information
-
The removal of the N-terminal hydrophobic sequence of NADPH-cytochrome P450 reductase results in loss of the ability of the flavoprotein to interact and transfer electrons to cytochrome P450. Truncated forms of the flavoprotein (residue 46-676 of the mutant (Q56Q) or 57-676 of the wild-type NADPH-cytochrome P450 reductase) are unable to transfer electrons to cytochrome P450c17 or P4503A4
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Kurzban, G.P.; Howarth, J.; Palmer, G.; Strobel, H.W.
NADPH-cytochrome P-450 reductase. Physical properties and redox behavior in the absence of the FAD moiety
J. Biol. Chem.
265
12272-12279
1990
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Vibet, A.; Dintinger, T.; Maboundou, J.C.; Gaillard, J.L.; Divoux, D.; Silberzahn, P.
Estrogen synthetase in the horse. Comparison of equine placental and rat liver NADPH-cytrochrome c (P-450) reductase activities
FEBS Lett.
261
31-34
1990
Equus caballus, Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Benveniste, I.; Lesot, A.; Hasenfratz, M.; Durst, F.
Immunochemical characterization of NADPH-cytochrome P-450 reductase from Jerusalem artichoke and other higher plants
Biochem. J.
259
847-853
1989
Saccharomyces cerevisiae, Helianthus tuberosus, Embryophyta, Locusta migratoria migratorioides, Rattus norvegicus, Sus scrofa
brenda
Ardies, C.M.; Lasker, J.M.; Bloswick, B.P.; Lieber, C.S.
Purification of NADPH:cytochrome c (cytochrome P-450) reductase from hamster liver microsomes by detergent extraction and affinity chromatography
Anal. Biochem.
162
39-46
1987
Mesocricetus auratus, Rattus norvegicus
brenda
Takahashi, N.; Saito, T.; Goda, Y.; Tomita, K.
Characterization of microsomal NADPH-dependent aldehyde reductase from rat brain
J. Biochem.
99
513-519
1986
Rattus norvegicus, Rattus norvegicus Wistar
brenda
Shephard, E.A.; Pike, S.F.; Rabin, B.R.; Phillips, I.R.
A rapid one-step purification of NADPH-cytochrome c (P-450) reductase from rat liver microsomes
Anal. Biochem.
129
430-433
1983
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Crankshaw, D.L.; Hetnarski, K.; Wilkinson, C.F.
Purification and characterization of NADPH--cytochrome c reductase from the midgut of the southern armyworm (Spodoptera eridania)
Biochem. J.
181
593-605
1979
Rattus norvegicus, Rattus norvegicus Sprague-Dawley, Spodoptera eridania
brenda
Zimmermann, J.J.; Kasper, C.B.
Immunological and biochemical characterization of nuclear envelope reduced nicotinamide adenine dinucleotide phosphate-cytochrome c oxidoreductase
Arch. Biochem. Biophys.
190
726-735
1978
Rattus norvegicus
brenda
Yasukochi, Y.; Masters, B.S.S.
Some properties of a detergent-solubilized NADPH-cytochrome c(cytochrome P-450) reductase purified by biospecific affinity chromatography
J. Biol. Chem.
251
5337-5344
1976
Rattus norvegicus, Sus scrofa
brenda
Masters, B.S.S.; Prough, R.A.; Kamin, H.
Properties of the stable aerobic and anaerobic half-reduced states of NADPH-cytochrome c reductase
Biochemistry
14
607-613
1975
Rattus norvegicus, Sus scrofa
brenda
Teschke, R.; Hasumura, Y.; Lieber, C.S.
Hepatic microsomal ethanol-oxidizing system: solubilization, isolation, and characterization
Arch. Biochem. Biophys.
163
404-415
1974
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Betz, G.; Roper, M.; Tsai, P.
Steroid 17,20-lyase from testis microsomes: participation of NADPH cytochrome c reductase
Arch. Biochem. Biophys.
163
318-323
1974
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Golf, S.W.; Graef, V.; Staudinger, H.
Solubilization and purification of the NADPH-cytochrome reductase from rat liver microsomes
Hoppe-Seyler's Z. Physiol. Chem.
355
1063-1069
1974
Rattus norvegicus, Rattus norvegicus Wistar
brenda
Omura, T.; Takesue, S.
A new method for simultaneous purification of cytochrome b5 and NADPH-cytochrome c reductase from rat liver microsomes
J. Biochem.
67
249-257
1970
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Cooper, M.B.; Craft, J.A.; Estall, M.R.; Rabin, B.R.
Asymmetric distribution of cytochrome P-450 and NADPH-cytochrome P-450 (cytochrome c) reductase in vesicles from smooth endoplasmic reticulum of rat liver
Biochem. J.
190
737-746
1980
Rattus norvegicus
brenda
Yasukochi, Y.; Masters, B.S.S.
Tetramer-dimer dissociation in homoglobin and the Bohr effect
J. Biol. Chem.
251
5537-5544
1976
Rattus norvegicus
brenda
Erecinska, M.
Cytochrome c Interaction with membranes, formylated cytochrome c1
Arch. Biochem. Biophys.
169
199-208
1975
Rattus norvegicus
brenda
Dialameh, G.H.
Isolation and partial purification of cytochrome-P-450 from induced rat liver
Biochem. Biophys. Res. Commun.
81
19-23
1978
Rattus norvegicus
brenda
Hara, T.; Taniguchi, M.
Relationship between changes in properties and contents of riboflavin derivatives of NADPH-cytochrome P-450 reductase in the liver microsomes of riboflavin-deficient rats
J. Biochem.
97
473-482
1985
Rattus norvegicus, Rattus norvegicus Wistar
brenda
Gut, J.; Kawato, S.; Cherry, R.J.; Winterhalter, K.H.; Richter, C.
Lipid peroxidation decreases the rotational mobility of cytochrome P-450 in rat liver microsomes
Biochim. Biophys. Acta
817
217-228
1985
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Lindeskog, P.; Haaparanta, T.; Norgard, M.; Glaumann, H.; Hansson, T.; Gustafsson J.
Isolation of rat intestinal microsomes: Partial characterisation of mucosal cytochrome P-450
Arch. Biochem. Biophys.
244
492-501
1986
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Balvers, W.G.; Boersma, M.G.; Vervoort, J.; Ouwehand, A.; Rietjens, I.M.C.M.
A specific interaction between NADPH-cytochrome reductase and phosphatidylserine and phosphatidylinositol
Eur. J. Biochem.
218
1021-1029
1993
Rattus norvegicus, Rattus norvegicus Wistar
brenda
McGuire, J.J.; Anderson, D.J.; McDonald, B.J.; Narayanasami, R.; Bennett, B.M.
Inhibition of NADPH-Cytochrome P450 Reductase and Glyceryl Trinitrate Biotransformation by Diphenyleneiodonium Sulfate
Biochem. Pharmacol.
56
881-893
1998
Rattus norvegicus, Rattus norvegicus Sprague-Dawley
brenda
Shen, A.L.; Porter, T.D.; Wilson, T.E.; Kasper, C.B.
Structural analysis of the FMN binding domain of NADPH-cytochrome P-450 oxidoreductase by site-directed mutagenesis
J. Biol. Chem.
264
7584-7589
1989
Rattus norvegicus
brenda
Shen, A.L.; Kasper, C.B.
Differential contributions of NADPH-Cytochrome P450 oxidoreductase FAD binding site residues to flavin binding and catalysis
J. Biol. Chem.
275
41087-41091
2000
Rattus norvegicus (P00388)
brenda
Kutty, R.K.; Maines, M.D.
Characterization of an NADH-dependent haem-degrading system in ox heart mitochondria
Biochem. J.
246
467-474
1987
Rattus norvegicus
brenda
Vatsis, K.P.; Coon, M.J.
Oxidative aldehyde deformylation catalyzed by NADPH-cytochrome P450 reductase and the flavoprotein domain of neuronal nitric oxide synthase
Biochem. Biophys. Res. Commun.
337
1107-1111
2005
Rattus norvegicus
brenda
Bonina, T.A.; Gilep, A.A.; Estabrook, R.W.; Usanov, S.A.
Engineering of proteolytically stable NADPH-cytochrome P450 reductase
Biochemistry
70
357-365
2005
Rattus norvegicus
brenda
Higashimoto, Y.; Sato, H.; Sakamoto, H.; Takahashi, K.; Palmer, G.; Noguchi, M.
The reactions of heme- and verdoheme-heme oxygenase-1 complexes with FMN-depleted NADPH-cytochrome P450 reductase. Electrons required for verdoheme oxidation can be transferred through a pathway not involving FMN
J. Biol. Chem.
281
31659-31667
2006
Rattus norvegicus
brenda
Yim, S.K.; Jung, H.C.; Pan, J.G.; Kang, H.S.; Ahn, T.; Yun, C.H.
Functional expression of mammalian NADPH-cytochrome P450 oxidoreductase on the cell surface of Escherichia coli
Protein Expr. Purif.
49
292-298
2006
Rattus norvegicus
brenda
Higashimoto, Y.; Sugishima, M.; Sato, H.; Sakamoto, H.; Fukuyama, K.; Palmer, G.; Noguchi, M.
Mass spectrometric identification of lysine residues of heme oxygenase-1 that are involved in its interaction with NADPH-cytochrome P450 reductase
Biochem. Biophys. Res. Commun.
367
852-858
2008
Rattus norvegicus (P00388)
brenda
Huber, W.J.; Backes, W.L.
Expression and characterization of full-length human heme oxygenase-1: the presence of intact membrane-binding region leads to increased binding affinity for NADPH cytochrome P450 reductase
Biochemistry
46
12212-12219
2007
Rattus norvegicus
brenda
Cherala, G.; Shapiro, B.H.; Dmello, A.P.
Effect of perinatal low protein diets on the ontogeny of select hepatic cytochrome p450 enzymes and cytochrome p450 reductase in the rat
Drug Metab. Dispos.
35
1057-1063
2007
Rattus norvegicus
brenda
Yao, H.T.; Chang, Y.W.; Lan, S.J.; Yeh, T.K.
The inhibitory effect of tannic acid on cytochrome P450 enzymes and NADPH-CYP reductase in rat and human liver microsomes
Food Chem. Toxicol.
46
645-653
2008
Homo sapiens, Rattus norvegicus
brenda
Wang, Y.; Gray, J.P.; Mishin, V.; Heck, D.E.; Laskin, D.L.; Laskin, J.D.
Role of cytochrome P450 reductase in nitrofurantoin-induced redox cycling and cytotoxicity
Free Radic. Biol. Med.
44
1169-1179
2008
Rattus norvegicus
brenda
Yim, S.K.; Jung, H.C.; Yun, C.H.; Pan, J.G.
Functional expression in Bacillus subtilis of mammalian NADPH-cytochrome P450 oxidoreductase and its spore-display
Protein Expr. Purif.
63
5-11
2008
Rattus norvegicus
brenda
Mast, N.; Liao, W.L.; Pikuleva, I.A.; Turko, I.V.
Combined use of mass spectrometry and heterologous expression for identification of membrane-interacting peptides in cytochrome P450 46A1 and NADPH-cytochrome P450 oxidoreductase
Arch. Biochem. Biophys.
483
81-89
2009
Rattus norvegicus (P00388)
brenda
Hall, C.N.; Keynes, R.G.; Garthwaite, J.
Cytochrome P450 oxidoreductase participates in nitric oxide consumption by rat brain
Biochem. J.
419
411-418
2009
Rattus norvegicus
brenda
Huber III, W.J.; Scruggs, B.A.; Backes, W.L.
C-Terminal membrane spanning region of human heme oxygenase-1 mediates a time-dependent complex formation with cytochrome P450 reductase
Biochemistry
48
190-197
2009
Rattus norvegicus
brenda
Kim, D.H.; Yim, S.K.; Kim, K.H.; Ahn, T.; Yun, C.H.
Continuous spectrofluorometric and spectrophotometric assays for NADPH-cytochrome P450 reductase activity using 5-cyano-2,3-ditolyl tetrazolium chloride
Biotechnol. Lett.
31
271-275
2009
Rattus norvegicus
brenda
Hamdane, D.; Xia, C.; Im, S.C.; Zhang, H.; Kim, J.J.; Waskell, L.
Structure and function of an NADPH-cytochrome P450 oxidoreductase in an open conformation capable of reducing cytochrome P450
J. Biol. Chem.
284
11374-11384
2009
Rattus norvegicus (P00388)
brenda
Guengerich, F.P.; Martin, M.V.; Sohl, C.D.; Cheng, Q.
Measurement of cytochrome P450 and NADPH-cytochrome P450 reductase
Nat. Protoc.
4
1245-1251
2009
Rattus norvegicus
brenda
Inaoka, Y.; Yazawa, T.; Mizutani, T.; Kokame, K.; Kangawa, K.; Uesaka, M.; Umezawa, A.; Miyamoto, K.
Regulation of P450 oxidoreductase by gonadotropins in rat ovary and its effect on estrogen production
Reprod. Biol. Endocrinol.
6
62
2008
Rattus norvegicus (P00388), Homo sapiens (P16435), Homo sapiens
brenda
Grinkova, Y.V.; Denisov, I.G.; Sligar, S.G.
Functional reconstitution of monomeric CYP3A4 with multiple cytochrome P450 reductase molecules in Nanodiscs
Biochem. Biophys. Res. Commun.
398
194-198
2010
Rattus norvegicus
brenda
Jang, H.H.; Jamakhandi, A.P.; Sullivan, S.Z.; Yun, C.H.; Hollenberg, P.F.; Miller, G.P.
Beta sheet 2-alpha helix C loop of cytochrome P450 reductase serves as a docking site for redox partners
Biochim. Biophys. Acta
1804
1285-1293
2010
Rattus norvegicus
brenda
Laursen, T.; Jensen, K.; Moller, B.L.
Conformational changes of the NADPH-dependent cytochrome P450 reductase in the course of electron transfer to cytochromes P450
Biochim. Biophys. Acta
1814
132-138
2011
Rattus norvegicus
brenda
Flueck, C.E.; Mullis, P.E.; Pandey, A.V.
Modeling of human P450 oxidoreductase structure by in silico mutagenesis and MD simulation
Mol. Cell. Endocrinol.
313
17-22
2009
Rattus norvegicus (P00388), Homo sapiens (P16435), Homo sapiens
brenda
Gray, J.P.; Mishin, V.; Heck, D.E.; Laskin, D.L.; Laskin, J.D.
Inhibition of NADPH cytochrome P450 reductase by the model sulfur mustard vesicant 2-chloroethyl ethyl sulfide is associated with increased production of reactive oxygen species
Toxicol. Appl. Pharmacol.
247
76-82
2010
Homo sapiens, Mus musculus, Rattus norvegicus
brenda
Iyanagi, T.; Xia, C.; Kim, J.J.
NADPH-cytochrome P450 oxidoreductase: prototypic member of the diflavin reductase family
Arch. Biochem. Biophys.
528
72-89
2012
Homo sapiens, Rattus norvegicus
brenda
Kenaan, C.; Zhang, H.; Shea, E.V.; Hollenberg, P.F.
Uncovering the role of hydrophobic residues in cytochrome P450-cytochrome P450 reductase interactions
Biochemistry
50
3957-3967
2011
Rattus norvegicus
brenda
Xia, C.; Hamdane, D.; Shen, A.L.; Choi, V.; Kasper, C.B.; Pearl, N.M.; Zhang, H.; Im, S.C.; Waskell, L.; Kim, J.J.
Conformational changes of NADPH-cytochrome P450 oxidoreductase are essential for catalysis and cofactor binding
J. Biol. Chem.
286
16246-16260
2011
Rattus norvegicus (P00388)
brenda
Shinkai, Y.; Nishihara, Y.; Amamiya, M.; Wakayama, T.; Li, S.; Kikuchi, T.; Nakai, Y.; Shimojo, N.; Kumagai, Y.
NADPH-cytochrome P450 reductase-mediated denitration reaction of 2,4,6-trinitrotoluene to yield nitrite in mammals
Free Radic. Biol. Med.
91
178-187
2016
Rattus norvegicus
brenda
Huang, R.; Zhang, M.; Rwere, F.; Waskell, L.; Ramamoorthy, A.
Kinetic and structural characterization of the interaction between the FMN binding domain of cytochrome P450 reductase and cytochrome c
J. Biol. Chem.
290
4843-4855
2015
Rattus norvegicus
brenda
Dai, Y.; Zhen, J.; Zhang, X.; Zhong, Y.; Liu, S.; Sun, Z.; Guo, Y.; Wu, Q.
Analysis of the complex formation, interaction and electron transfer pathway between the open conformation of NADPH-cytochrome P450 reductase and aromatase
Steroids
101
116-124
2015
Rattus norvegicus (P00388)
brenda
Nemeikaite-Ceniene, A.; Sarlauskas, J.; Miseviciene, L.; Maroziene, A.; Jonusiene, V.; Lesanavicius, M.; Cenas, N.
Aerobic cytotoxicity of aromatic N-oxides the role of NAD(P)H quinone oxidoreductase (NQO1)
Int. J. Mol. Sci.
21
8754
2020
Rattus norvegicus (P00388)
brenda
Park, H.G.; Lim, Y.R.; Han, S.; Jeong, D.; Kim, D.
Enhanced purification of recombinant rat NADPH-P450 reductase by using a hexahistidine-tag
J. Microbiol. Biotechnol.
27
983-989
2017
Rattus norvegicus (P00388)
brenda