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Literature summary extracted from

  • Shen, A.L.; Kasper, C.B.
    Differential contributions of NADPH-Cytochrome P450 oxidoreductase FAD binding site residues to flavin binding and catalysis (2000), J. Biol. Chem., 275, 41087-41091.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.6.2.4 expression in Escherichia coli Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
1.6.2.4 C472T substitution does not affect FAD or FMN incorporation, substitution has no effect on activity, Km for NADPH or Km for cytochrome c Rattus norvegicus
1.6.2.4 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 Rattus norvegicus
1.6.2.4 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 Rattus norvegicus
1.6.2.4 S678X substitution does not affect FAD or FMN incorporation, substitution has no effect on the catalytic activity or kinetic properties Rattus norvegicus
1.6.2.4 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 Rattus norvegicus
1.6.2.4 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 Rattus norvegicus
1.6.2.4 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 Rattus norvegicus
1.6.2.4 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 Rattus norvegicus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.6.2.4 0.000001
-
 NADPH wild type, cosubstrate FAD, Km below Rattus norvegicus
1.6.2.4 0.000001
-
 cytochrome c wild type, cosubstrate FAD, Km below Rattus norvegicus
1.6.2.4 0.00012
-
 NADPH T491V mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.00012
-
 cytochrome c T491V mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0008
-
 NADPH G488L mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0008
-
 cytochrome c G488L mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0015
-
 cytochrome c W677X mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0028
-
 cytochrome c W677Y mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0041
-
 cytochrome c R454E mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0056
-
 cytochrome c S678X mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0058
-
 cytochrome c Y456S mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0059
-
 cytochrome c T491V mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0062
-
 cytochrome c wild type, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0063
-
 cytochrome c C472T mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0077
-
 cytochrome c G488L mutant, cosubstrate NADPH Rattus norvegicus
1.6.2.4 0.0078
-
 NADPH Y456S mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0078
-
 cytochrome c Y456S mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0085
-
 NADPH W677X mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0134
-
 NADPH G488L mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0143
-
 NADPH W677Y mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0163
-
 NADPH wild type, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0166
-
 NADPH R454E mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0199
-
 NADPH S678X mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0201
-
 NADPH T491V mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0217
-
 NADPH C472T mutant, cosubstrate cytochrome c Rattus norvegicus
1.6.2.4 0.0251
-
 NADPH R454E mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0251
-
 cytochrome c R454E mutant, cosubstrate FAD Rattus norvegicus
1.6.2.4 0.0548
-
 NADPH Y456S mutant, cosubstrate cytochrome c Rattus norvegicus

Organism

EC Number Organism UniProt Comment Textmining
1.6.2.4 Rattus norvegicus P00388
-
-

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.6.2.4 liver
-
 Rattus norvegicus
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.6.2.4 0.17
-
 R454E mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 0.23
-
 Y456S mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 1.35
-
 G488L mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 1.7
-
 W677X mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 32.7
-
 T491V mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 36
-
 W677Y mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 57.4
-
 wild type, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 62.5
-
 S678X mutant, electron acceptor: cytochrome c Rattus norvegicus
1.6.2.4 65.4
-
 C472T mutant, electron acceptor: cytochrome c Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.2.4 2 ferricytochrome c + NADPH
-
 Rattus norvegicus 2 ferrocytochrome c + NADP+ + H+
-
 ?

Cofactor

EC Number Cofactor Comment Organism Structure
1.6.2.4 FAD
-
 Rattus norvegicus
1.6.2.4 FMN
-
 Rattus norvegicus
1.6.2.4 NADPH
-
 Rattus norvegicus