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Literature summary for 1.6.2.4 extracted from

  • Meints, C.E.; Parke, S.M.; Wolthers, K.R.
    Proximal FAD histidine residue influences interflavin electron transfer in cytochrome P450 reductase and methionine synthase reductase (2014), Arch. Biochem. Biophys., 547, 18-26 .
    View publication on PubMed
Search for more literature for 1.6.2.4

Protein Variants

Protein Variants Comment Organism
H322Ala the mutation does not affect the rate of NADPH hydride transfer or the FAD redox potentials, but does impede interflavin electron transfer. The mutant elicits a 4fold decrease in cytochrome c reduction and a 1.5fold decrease in ferricyanide reduction. The H322A substitution also leads to a modest increase in NADP(H) binding affinity, evidenced by a 2-3fold reduction in the Km for NADPH and Ki for NADP+ Homo sapiens
H322Q the mutant shows a 50% decrease in cytochrome c and ferricyanide reduction and a marginal increase in NADP(H) binding affinity compared to the wild type enzyme Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
NADP+
-
 Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00031
-
 NADPH mutant enzyme H322A, at pH 7.5 and 25°C Homo sapiens
0.00058
-
 NADPH mutant enzyme H322Q, at pH 7.5 and 25°C Homo sapiens
0.00071
-
 NADPH wild type enzyme, at pH 7.5 and 25°C Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NADPH + H+ + ferricyanide
-
 Homo sapiens NADP+ + ferrocyanide
-
 ?
NADPH + H+ + ferricytochrome c
-
 Homo sapiens NADP+ + ferrocytochrome c
-
 ?

Synonyms

Synonyms Comment Organism
CPR
-
 Homo sapiens
cytochrome P450 reductase
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.4
-
 NADPH mutant enzyme H322A, at pH 7.5 and 25°C Homo sapiens
11.3
-
 NADPH mutant enzyme H322Q, at pH 7.5 and 25°C Homo sapiens
18.1
-
 ferricyanide mutant enzyme H322A, at pH 7.5 and 25°C Homo sapiens
20
-
 NADPH wild type enzyme, at pH 7.5 and 25°C Homo sapiens
26.1
-
 ferricyanide mutant enzyme H322Q, at pH 7.5 and 25°C Homo sapiens
43.5
-
 ferricyanide wild type enzyme, at pH 7.5 and 25°C Homo sapiens

Cofactor

Cofactor Comment Organism Structure
FAD
-
 Homo sapiens
FMN
-
 Homo sapiens

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0003
-
 NADP+ mutant enzyme H322A, at pH 7.5 and 25°C Homo sapiens
0.00065
-
 NADP+ mutant enzyme H322Q, at pH 7.5 and 25°C Homo sapiens
0.00095
-
 NADP+ wild type enzyme, at pH 7.5 and 25°C Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
17400
-
 NADPH mutant enzyme H322A, at pH 7.5 and 25°C Homo sapiens
19400
-
 NADPH mutant enzyme H322Q, at pH 7.5 and 25°C Homo sapiens
28200
-
 NADPH wild type enzyme, at pH 7.5 and 25°C Homo sapiens