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Literature summary extracted from

  • Faro, M.; Frago, S.; Mayoral, T.; Hermoso, J.A.; Sanz-Aparicio, J.; Gomez-Moreno, C.; Medina, M.
    Probing the role of glutamic acid 139 of Anabaena ferredoxin-NADP+ reductase in the interaction with substrates (2002), Eur. J. Biochem., 269, 4938-4947.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.18.1.2 expression of wild-type and mutant enzymes Anabaena sp.

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.18.1.2 purified recombinant mutant E139K, hanging drop vapour diffusion method, 0.002 ml protein solution containing 0.75 mM protein in 10 mM Tris-HCl, pH 8.0, plus 0.001 ml reservoir solution containing 5% w/v beta-octylglycoside, 18% PEG 6000, 20 mM ammonium sulfate, and 0.1 M MES/NaOH, pH 5.5, equilibration against 1 ml reservoir solution at 20°C, phase separation due to detergent, X-ray diffraction structure determination and analysis at 2.5 A resolution Anabaena sp.

Protein Variants

EC Number Protein Variants Comment Organism
1.18.1.2 E139D site-directed mutagenesis, altered conformation compared to the wild-type enzyme, slightly reduced activity compared to the wild-type enzyme Anabaena sp.
1.18.1.2 E139K site-directed mutagenesis, mutant enzyme shows increased interaction with ferredoxin and reduces the appropriate orientation of flavodoxin, altered conformation compared to the wild-type enzyme, increased activity compared to the wild-type enzyme Anabaena sp.
1.18.1.2 E139Q site-directed mutagenesis, mutant enzyme shows increased interaction with ferredoxin and reduces the appropriate orientation of flavodoxin, altered conformation compared to the wild-type enzyme, increased activity compared to the wild-type enzyme Anabaena sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.18.1.2 additional information activity decreases with elevated ionic strength Anabaena sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.18.1.2 additional information
-
 additional information stopped-flow kinetics measurements, pH 8.0, 13°C, steady-state kinetics for wild-type and mutant enzymes dependent on ionic strength, overview Anabaena sp.
1.18.1.2 0.00027
-
 oxidized ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.0005
-
 oxidized ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 0.0009
-
 oxidized ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 0.0025
-
 oxidized ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 0.0034
-
 NADPH recombinant mutant E139Q, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.0035
-
 oxidized ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 0.0043
-
 oxidized ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.0047
-
 NADPH recombinant mutant E139D, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.0058
-
 NADPH recombinant mutant E139K, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.006
-
 NADPH recombinant wild-type enzyme, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.02
-
 oxidized ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 0.021
-
 oxidized ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 0.023
-
 oxidized ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 0.023
-
 oxidized ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.1
-
 oxidized ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 0.4
-
 oxidized ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.18.1.2 reduced ferredoxin + NADP+ Anabaena sp.
-
 oxidized ferredoxin + NADPH
-
 r

Organism

EC Number Organism UniProt Comment Textmining
1.18.1.2 Anabaena sp.
-
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
1.18.1.2 2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH Glu139 is essential for interaction with the substrate steering ferredoxin, flavodoxin and NADP+/NADPH in appropriate position for docking, Glu139 is not involved in binding Anabaena sp.
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.18.1.2 additional information NADPH-dependent cytochrome c reductase assay for determination of activity with ferredoxin or flavodoxin as electron carrier Anabaena sp. ?
-
 ?
1.18.1.2 oxidized 2,6-dichlorophenolindophenol + NADP+ diaphorase activity Anabaena sp. reduced 2,6-dichlorophenolindophenol + NADPH
-
 ?
1.18.1.2 oxidized ferredoxin + NADPH + H+
-
 Anabaena sp. reduced ferredoxin + NADP+
-
 r
1.18.1.2 reduced ferredoxin + NADP+
-
 Anabaena sp. oxidized ferredoxin + NADPH
-
 r
1.18.1.2 reduced ferredoxin + NADP+ hydride transfer of the N5 of the FAD isoalloxazine ring to the NADP+ nicotinamide ring, transfer of 2 electrons via the one-electron-carrier ferredoxin Anabaena sp. oxidized ferredoxin + NADPH
-
 r
1.18.1.2 reduced ferredoxin + NADP+
-
 Anabaena sp. oxidized ferredoxin + NADPH + H+
-
 r
1.18.1.2 reduced flavodoxin + NADP+
-
 Anabaena sp. oxidized flavodoxin + NADPH + H+
-
 r

Subunits

EC Number Subunits Comment Organism
1.18.1.2 More three-dimensional structure of mutant E139K Anabaena sp.

Synonyms

EC Number Synonyms Comment Organism
1.18.1.2 ferredoxin-NADP+ reductase
-
 Anabaena sp.
1.18.1.2 FNR
-
 Anabaena sp.

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.18.1.2 25
-
 assay at Anabaena sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.18.1.2 2 8 reduced flavodoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 14
-
 reduced flavodoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 17
-
 reduced flavodoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 19
-
 reduced flavodoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 20
-
 reduced flavodoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 24
-
 reduced flavodoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 25
-
 reduced flavodoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 25
-
 reduced flavodoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 26
-
 reduced flavodoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 38
-
 reduced flavodoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 58
-
 reduced ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 59
-
 NADPH recombinant mutant E139K, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 70
-
 reduced ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 81
-
 NADPH recombinant wild-type enzyme, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 88
-
 NADPH recombinant mutant E139Q, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 89
-
 NADPH recombinant mutant E139D, diaphorase activity with 2,6-dichlorophenolindophenol, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 117
-
 reduced ferredoxin recombinant mutant E139Q, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 120
-
 reduced ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 135
-
 reduced ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 148
-
 reduced ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 200 mM Anabaena sp.
1.18.1.2 155
-
 reduced ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 176
-
 reduced ferredoxin recombinant mutant E139K, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 192
-
 reduced ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 209
-
 reduced ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 100 mM Anabaena sp.
1.18.1.2 225
-
 reduced ferredoxin recombinant wild-type enzyme, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.
1.18.1.2 280
-
 reduced ferredoxin recombinant mutant E139D, pH 8.0, 25°C, ionic strength of 28 mM Anabaena sp.

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.18.1.2 8
-
 assay at Anabaena sp.

Cofactor

EC Number Cofactor Comment Organism Structure
1.18.1.2 FAD flavoenzyme Anabaena sp.