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Literature summary for 1.6.2.4 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
Search for more literature for 1.6.2.4

Cloned(Commentary)

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

Inhibitors

Inhibitors Comment Organism Structure
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]

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

Metals/Ions

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

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

Organism

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

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.15
-
 isozyme CPR2, using ferricyanide as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
3.17
-
 isozyme CPR2, using ferricytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
3.3
-
 isozyme CPR2, for NADPH with cytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
3.47
-
 isozyme CPR1, using ferricyanide as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
4.69
-
 isozyme CPR1, using ferricytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
4.78
-
 isozyme CPR1, for NADPH with cytochrome c as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
9.32
-
 isozyme CPR1, using dichlorophenolindophenol as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum
9.84
-
 isozyme CPR2, using dichlorophenolindophenol as cosubstrate, at 25°C, pH 7.4 Gossypium hirsutum

Substrates and Products (Substrate)

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

Synonyms

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

Turnover Number [1/s]

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

Cofactor

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

Expression

Organism Comment Expression
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
Gossypium hirsutum isozyme CPR2 shows a dramatic increase of transcript abundance around 15-20 days post anthesis up

General Information

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

kcat/KM [mM/s]

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