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Literature summary extracted from

  • Yang, C.Q.; Lu, S.; Mao, Y.B.; Wang, L.J.; Chen, X.Y.
    Characterization of two NADPH: cytochrome P450 reductases from cotton (Gossypium hirsutum) (2010), Phytochemistry, 71, 27-35.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.6.2.4 expressed in Escherichia coli BL21(DE3) cells Gossypium hirsutum

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.6.2.4 diphenyleneiodonium about 60% inhibition at 0.02 mM and more than 90% inhibition at 0.06 mM; about 60% inhibition at 0.02 mM and more than 90% inhibition at 0.06 mM Gossypium hirsutum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.6.2.4 0.0012
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 0.0016
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 0.0046
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 0.0056
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.6.2.4 K+ isozyme CPR1 increases to about 2old in the presence of 0.1 M KCl, then decreases at higher concentrations of KCl Gossypium hirsutum
1.6.2.4 K+ isozyme CPR2 increases to about 2old in the presence of 0.1 M KCl, then decreases at higher concentrations of KCl Gossypium hirsutum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.6.2.4 77140
-
isozyme CPR1, calculated from amino acid sequence Gossypium hirsutum
1.6.2.4 78590
-
isozyme CPR2, calculated from amino acid sequence Gossypium hirsutum

Organism

EC Number Organism UniProt Comment Textmining
1.6.2.4 Gossypium hirsutum C0LLU4 cultivars CCRI1 2 and CCRI20
-
1.6.2.4 Gossypium hirsutum C0LLU5 cultivars CCRI1 2 and CCRI20
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.6.2.4 Ni-NTA resin column chromatography Gossypium hirsutum

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 2.15
-
isozyme CPR2, using ferricyanide as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 3.17
-
isozyme CPR2, using ferricytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 3.3
-
isozyme CPR2, for NADPH with cytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 3.47
-
isozyme CPR1, using ferricyanide as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 4.69
-
isozyme CPR1, using ferricytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 4.78
-
isozyme CPR1, for NADPH with cytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 9.32
-
isozyme CPR1, using dichlorophenolindophenol as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 9.84
-
isozyme CPR2, using dichlorophenolindophenol as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.6.2.4 2 ferricyanide + NADPH
-
Gossypium hirsutum 2 ferrocyanide + NADP+ + H+
-
?
1.6.2.4 2 ferricytochrome c + NADPH
-
Gossypium hirsutum 2 ferrocytochrome c + NADP+ + H+
-
?
1.6.2.4 2 ferricytochrome P450 + NADPH
-
Gossypium hirsutum 2 ferrocytochrome P450 + NADP+ + H+
-
?
1.6.2.4 additional information no activity with NADH Gossypium hirsutum ?
-
?
1.6.2.4 NADPH + H+ + dichlorophenolindophenol
-
Gossypium hirsutum NADP+ + reduced dichlorophenolindophenol
-
?

Synonyms

EC Number Synonyms Comment Organism
1.6.2.4 CPR1 isozyme Gossypium hirsutum
1.6.2.4 CPR2 isozyme Gossypium hirsutum
1.6.2.4 cytochrome P450 reductase
-
Gossypium hirsutum
1.6.2.4 NADPH: cytochrome P450 reductase
-
Gossypium hirsutum

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.6.2.4 3.86
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 3.97
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 10.1
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 10.2
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum

Cofactor

EC Number Cofactor Comment Organism Structure
1.6.2.4 FAD one mol isozyme CPR1 contains 0.87 mol of FAD Gossypium hirsutum
1.6.2.4 FAD one mole isozyme CPR2 contains 0.91 mol of FAD Gossypium hirsutum
1.6.2.4 FMN one mol isozyme CPR1 contains 0.83 mol of FMN Gossypium hirsutum
1.6.2.4 FMN one mole isozyme CPR2 contains 0.86 mol of FMN Gossypium hirsutum
1.6.2.4 NADPH
-
Gossypium hirsutum

Expression

EC Number Organism Comment Expression
1.6.2.4 Gossypium hirsutum expression of isozyme CPR2, but not CPR1, is inducible by mechanical wounding and elicitation. Isozyme CPR1 is expressed at a relatively constant level throughout the developmental stages investigated (0-40 days post anthesis), with a slight up-regulation at 15 days post anthesis up
1.6.2.4 Gossypium hirsutum isozyme CPR2 shows a dramatic increase of transcript abundance around 15-20 days post anthesis up

General Information

EC Number General Information Comment Organism
1.6.2.4 physiological function the recombinant enzymes supports the activity of CYP73A25, a cinnamate 4-hydroxylase of cotton Gossypium hirsutum

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.6.2.4 838
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 1825
-
NADPH isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 3321
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum
1.6.2.4 6305
-
ferricytochrome c isozyme CPR1, at 25°C, pH 7.4 Gossypium hirsutum