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Literature summary for 1.6.2.4 extracted from

  • 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 (1989), J. Biol. Chem., 264, 7584-7589.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
Y140D substitution does not eliminate FMN binding but reduces cytochrome c reductase activity, Km value for cytochrome c or NADPH similar to wild type Rattus norvegicus
Y140D/178D substitution abolishes FMN binding and cytochrome c reductase activity Rattus norvegicus
Y140F substitution has no effect on FMN content or catalytic activity, Km value for cytochrome c or NADPH similar to wild type Rattus norvegicus
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 Rattus norvegicus
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 Rattus norvegicus
Y178F substitution has no effect on FMN content or catalytic activity, slightly decreases Km for cytochrome c, NADPH Km value similar to wild type Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0064
-
NADPH wild type, cosubstrate cytochrome c Rattus norvegicus
0.0066
-
NADPH native enzyme, liver, cosubstrate cytochrome c Rattus norvegicus
0.0066
-
NADPH Y140D mutant, cosubstrate cytochrome c Rattus norvegicus
0.0071
-
NADPH Y178F mutant, cosubstrate cytochrome c Rattus norvegicus
0.0078
-
NADPH Y140F mutant, cosubstrate cytochrome c Rattus norvegicus
0.0095
-
NADPH Y178D mutant, cosubstrate cytochrome c Rattus norvegicus
0.0114
-
NADPH Y140F/178F double mutant, cosubstrate cytochrome c Rattus norvegicus
0.0131
-
cytochrome c Y140F/178F double mutant Rattus norvegicus
0.014
-
cytochrome c Y178F mutant Rattus norvegicus
0.0178
-
cytochrome c native enzyme, liver Rattus norvegicus
0.0185
-
cytochrome c Y178D mutant Rattus norvegicus
0.0188
-
cytochrome c Y140F mutant Rattus norvegicus
0.0191
-
cytochrome c Y140D mutant Rattus norvegicus
0.0211
-
cytochrome c wild type Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
80000
-
wild type, x * 80000, SDS-PAGE Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
expressed in Escherichia coli Rattus norvegicus

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
Rattus norvegicus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.28
-
Y140D/178D double mutant, electron acceptor: cytochrome c Rattus norvegicus
0.44
-
Y178D mutant, electron acceptor: cytochrome c Rattus norvegicus
11
-
Y140D mutant, electron acceptor: cytochrome c Rattus norvegicus
46
-
Y140F/178F double mutant, electron acceptor: cytochrome c Rattus norvegicus
47.4
-
Y178F mutant, electron acceptor: cytochrome c Rattus norvegicus
48.1
-
Y178D mutant, electron acceptor: ferricyanide Rattus norvegicus
51
-
Y140D/178D double mutant, electron acceptor: ferricyanide Rattus norvegicus
51.5
-
bacterially expressed reductase protein, electron acceptor: cytochrome c Rattus norvegicus
52.9
-
electron acceptor: ferricyanide Rattus norvegicus
53.3
-
electron acceptor: cytochrome c Rattus norvegicus
55.2
-
Y140F mutant, electron acceptor: cytochrome c Rattus norvegicus
94.2
-
Y140F/178F double mutant, electron acceptor: ferricyanide Rattus norvegicus
94.8
-
Y140F mutant, electron acceptor: ferricyanide Rattus norvegicus
102
-
bacterially expressed reductase protein, electron acceptor: ferricyanide Rattus norvegicus
102
-
Y140D mutant, electron acceptor: ferricyanide Rattus norvegicus
113
-
Y178F mutant, electron acceptor: ferricyanide Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2 ferricytochrome c + NADPH additional electron acceptor: ferricyanide Rattus norvegicus 2 ferrocytochrome c + NADP+ + H+
-
?

Subunits

Subunits Comment Organism
? wild type, x * 80000, SDS-PAGE Rattus norvegicus

Cofactor

Cofactor Comment Organism Structure
FAD
-
Rattus norvegicus
FMN
-
Rattus norvegicus