BRENDA - Enzyme Database

Two functionally distinct NADP+-dependent ferredoxin oxidoreductases maintain the primary redox balance of Pyrococcus furiosus

Nguyen, D.M.N.; Schut, G.J.; Zadvornyy, O.A.; Tokmina-Lukaszewska, M.; Poudel, S.; Lipscomb, G.L.; Adams, L.A.; Dinsmore, J.T.; Nixon, W.J.; Boyd, E.S.; Bothner, B.; Peters, J.W.; Adams, M.W.W.; J. Biol. Chem. 292, 14603-14616 (2017)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
1.18.1.2
vapor diffusion method, using 0.22 M magnesium sulfate, 27% (w/v) polyethylene glycol 3350, and 0.4% (v/v) ethyl acetate in the presence of 1 mM sodium dithionite
Pyrococcus furiosus
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.18.1.2
88951
-
mass spectrometry
Pyrococcus furiosus
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.18.1.2
reduced ferredoxin + NADP+ + H+
Pyrococcus furiosus
-
oxidized ferredoxin + NADPH
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
Pyrococcus furiosus COM1
-
oxidized ferredoxin + NADPH
-
-
?
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.6.1.4
Pyrococcus furiosus
Q8U195 and Q8U194
Q8U195: subunit alpha, Q8U194: subunit beta
-
1.18.1.2
Pyrococcus furiosus
I6TYZ3
-
-
1.18.1.2
Pyrococcus furiosus COM1
I6TYZ3
-
-
Purification (Commentary)
EC Number
Commentary
Organism
1.18.1.2
His-Trap column chromatography, Ni-NTA column chromatography, and S100 gel filtration
Pyrococcus furiosus
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.18.1.2
17
-
after 1fold purification, at pH 7.5 and 80░C
Pyrococcus furiosus
1.18.1.2
275
-
after 15fold purification, at pH 7.5 and 80░C
Pyrococcus furiosus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.1.4
NADH + H+ + 2 NADP+ + 2 reduced ferredoxin [iron-sulfur] cluster
-
745371
Pyrococcus furiosus
NAD+ + 2 NADPH + 2 oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.18.1.2
additional information
the enzyme does not perform NADH-dependent benzyl viologen reduction
745371
Pyrococcus furiosus
?
-
-
-
-
1.18.1.2
additional information
the enzyme does not perform NADH-dependent benzyl viologen reduction
745371
Pyrococcus furiosus COM1
?
-
-
-
-
1.18.1.2
reduced benzyl viologen + NADP+ + H+
-
745371
Pyrococcus furiosus
oxidized benzyl viologen + NADPH
-
-
-
?
1.18.1.2
reduced benzyl viologen + NADP+ + H+
-
745371
Pyrococcus furiosus COM1
oxidized benzyl viologen + NADPH
-
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
-
745371
Pyrococcus furiosus
oxidized ferredoxin + NADPH
-
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
-
745371
Pyrococcus furiosus COM1
oxidized ferredoxin + NADPH
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.18.1.2
heterodimer
1 * 52498 + 1 * 34029, calculated from amino acid sequence
Pyrococcus furiosus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.6.1.4
flavin
flavoprotein
Pyrococcus furiosus
1.18.1.2
FAD
-
Pyrococcus furiosus
1.18.1.2
NADP+
-
Pyrococcus furiosus
1.18.1.2
[2Fe-2S]-center
-
Pyrococcus furiosus
1.18.1.2
[4Fe-4S]-center
-
Pyrococcus furiosus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.6.1.4
flavin
flavoprotein
Pyrococcus furiosus
1.18.1.2
FAD
-
Pyrococcus furiosus
1.18.1.2
NADP+
-
Pyrococcus furiosus
1.18.1.2
[2Fe-2S]-center
-
Pyrococcus furiosus
1.18.1.2
[4Fe-4S]-center
-
Pyrococcus furiosus
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.18.1.2
vapor diffusion method, using 0.22 M magnesium sulfate, 27% (w/v) polyethylene glycol 3350, and 0.4% (v/v) ethyl acetate in the presence of 1 mM sodium dithionite
Pyrococcus furiosus
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.18.1.2
88951
-
mass spectrometry
Pyrococcus furiosus
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
1.18.1.2
reduced ferredoxin + NADP+ + H+
Pyrococcus furiosus
-
oxidized ferredoxin + NADPH
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
Pyrococcus furiosus COM1
-
oxidized ferredoxin + NADPH
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.18.1.2
His-Trap column chromatography, Ni-NTA column chromatography, and S100 gel filtration
Pyrococcus furiosus
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [Ámol/min/mg]
Specific Activity Maximum [Ámol/min/mg]
Commentary
Organism
1.18.1.2
17
-
after 1fold purification, at pH 7.5 and 80░C
Pyrococcus furiosus
1.18.1.2
275
-
after 15fold purification, at pH 7.5 and 80░C
Pyrococcus furiosus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.6.1.4
NADH + H+ + 2 NADP+ + 2 reduced ferredoxin [iron-sulfur] cluster
-
745371
Pyrococcus furiosus
NAD+ + 2 NADPH + 2 oxidized ferredoxin [iron-sulfur] cluster
-
-
-
?
1.18.1.2
additional information
the enzyme does not perform NADH-dependent benzyl viologen reduction
745371
Pyrococcus furiosus
?
-
-
-
-
1.18.1.2
additional information
the enzyme does not perform NADH-dependent benzyl viologen reduction
745371
Pyrococcus furiosus COM1
?
-
-
-
-
1.18.1.2
reduced benzyl viologen + NADP+ + H+
-
745371
Pyrococcus furiosus
oxidized benzyl viologen + NADPH
-
-
-
?
1.18.1.2
reduced benzyl viologen + NADP+ + H+
-
745371
Pyrococcus furiosus COM1
oxidized benzyl viologen + NADPH
-
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
-
745371
Pyrococcus furiosus
oxidized ferredoxin + NADPH
-
-
-
?
1.18.1.2
reduced ferredoxin + NADP+ + H+
-
745371
Pyrococcus furiosus COM1
oxidized ferredoxin + NADPH
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.18.1.2
heterodimer
1 * 52498 + 1 * 34029, calculated from amino acid sequence
Pyrococcus furiosus
General Information
EC Number
General Information
Commentary
Organism
1.6.1.4
metabolism
the enzyme has a key role in maintaining redox homeostasis. It is involved in maintaining the cellular redox balance, producing NADPH for biosynthesis by recycling the two other primary redox carriers, NADH and ferredoxin. The enzyme effectively couples the endergonic reduction of NADP+ by NADH and the exergonic reduction of NADP+ by reduced ferredoxin, thereby maintaining a high ratio of NADPH/NADP+ to drive biosynthesis. The Pyrococcus furiosus genome encodes an NfnI paralog termed NfnII, and the two are differentially expressed, depending on the growth conditions. Deletion of the genes encoding either NfnI or NfnII affects the cellular concentrations of NAD(P)H and particularly NADPH. This results in a moderate to severe growth phenotype in deletion mutants, demonstrating a key role for each enzyme in maintaining redox homeostasis
Pyrococcus furiosus
1.18.1.2
physiological function
the enzyme is important for growth in the presence of sulfur
Pyrococcus furiosus
General Information (protein specific)
EC Number
General Information
Commentary
Organism
1.6.1.4
metabolism
the enzyme has a key role in maintaining redox homeostasis. It is involved in maintaining the cellular redox balance, producing NADPH for biosynthesis by recycling the two other primary redox carriers, NADH and ferredoxin. The enzyme effectively couples the endergonic reduction of NADP+ by NADH and the exergonic reduction of NADP+ by reduced ferredoxin, thereby maintaining a high ratio of NADPH/NADP+ to drive biosynthesis. The Pyrococcus furiosus genome encodes an NfnI paralog termed NfnII, and the two are differentially expressed, depending on the growth conditions. Deletion of the genes encoding either NfnI or NfnII affects the cellular concentrations of NAD(P)H and particularly NADPH. This results in a moderate to severe growth phenotype in deletion mutants, demonstrating a key role for each enzyme in maintaining redox homeostasis
Pyrococcus furiosus
1.18.1.2
physiological function
the enzyme is important for growth in the presence of sulfur
Pyrococcus furiosus