BRENDA - Enzyme Database
show all sequences of 1.6.1.4

Mechanistic insights into energy conservation by flavin-based electron bifurcation

Lubner, C.E.; Jennings, D.P.; Mulder, D.W.; Schut, G.J.; Zadvornyy, O.A.; Hoben, J.P.; Tokmina-Lukaszewska, M.; Berry, L.; Nguyen, D.M.; Lipscomb, G.L.; Bothner, B.; Jones, A.K.; Miller, A.F.; King, P.W.; Adams, M.W.W.; Peters, J.W.; Nat. Chem. Biol. 13, 655-659 (2017)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
-
Pyrococcus furiosus
Organism
Organism
UniProt
Commentary
Textmining
Pyrococcus furiosus
Q8U195 and Q8U194
Q8U195: subunit alpha, Q8U194: subunit beta
-
Purification (Commentary)
Purification (Commentary)
Organism
-
Pyrococcus furiosus
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
NAD+ + 2 NADPH + 2 oxidized ferredoxin [iron-sulfur] cluster
reduction of both NAD+ and Fd by Nfn enzymes is coupled to the bifurcation of electrons from NADPH oxidation. The exergonic coupling of the NADP(H) and NAD(H) half reactions in Nfn is presumed to be mediated by NADPH oxidation at L-FAD followed by electron transfer to the proximal [2Fe-2S] cluster and subsequently the NAD(H) binding site at S-FAD. Biophysical analysis of the enzyme provides the first direct insight into the mechanism of flavin-based electron bifurcation and the requisite structural features
745843
Pyrococcus furiosus
NADH + H+ + 2 NADP+ + 2 reduced ferredoxin [iron-sulfur] cluster
-
-
-
?
Subunits
Subunits
Commentary
Organism
heterodimer
1 * 31000 + 1 * 53000, SDS-PAGE
Pyrococcus furiosus
Synonyms
Synonyms
Commentary
Organism
NfnI
-
Pyrococcus furiosus
Cofactor
Cofactor
Commentary
Organism
Structure
FAD
the 31000 Da subunit contains one FAD (S-FAD) and a [2Fe-2S] cluster with an unusual Asp ligand. The 53000 Da subunit contains one FAD (L-FAD), which is the site of electron bifurcation, and two [4Fe-4S] clusters. The L-FAD proximal [4Fe-4S] cluster coordination includes an unusual Glu ligand. S-FAD and L-FAD bind NADH and NADPH, respectively
Pyrococcus furiosus
[2Fe-2S]-center
the 31000 Da subunit contains one FAD (S-FAD) and a [2Fe-2S] cluster. The [2Fe-2S] cluster is unusual, with a relatively high (positive) reduction potential and coordination by one Asp and three Cys ligands
Pyrococcus furiosus
[4Fe-4S]-center
the 53000 Da subunit contains one FAD (L-FAD), which is the site of electron bifurcation, and two [4Fe-4S] clusters
Pyrococcus furiosus
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Pyrococcus furiosus
Cofactor (protein specific)
Cofactor
Commentary
Organism
Structure
FAD
the 31000 Da subunit contains one FAD (S-FAD) and a [2Fe-2S] cluster with an unusual Asp ligand. The 53000 Da subunit contains one FAD (L-FAD), which is the site of electron bifurcation, and two [4Fe-4S] clusters. The L-FAD proximal [4Fe-4S] cluster coordination includes an unusual Glu ligand. S-FAD and L-FAD bind NADH and NADPH, respectively
Pyrococcus furiosus
[2Fe-2S]-center
the 31000 Da subunit contains one FAD (S-FAD) and a [2Fe-2S] cluster. The [2Fe-2S] cluster is unusual, with a relatively high (positive) reduction potential and coordination by one Asp and three Cys ligands
Pyrococcus furiosus
[4Fe-4S]-center
the 53000 Da subunit contains one FAD (L-FAD), which is the site of electron bifurcation, and two [4Fe-4S] clusters
Pyrococcus furiosus
Purification (Commentary) (protein specific)
Commentary
Organism
-
Pyrococcus furiosus
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
NAD+ + 2 NADPH + 2 oxidized ferredoxin [iron-sulfur] cluster
reduction of both NAD+ and Fd by Nfn enzymes is coupled to the bifurcation of electrons from NADPH oxidation. The exergonic coupling of the NADP(H) and NAD(H) half reactions in Nfn is presumed to be mediated by NADPH oxidation at L-FAD followed by electron transfer to the proximal [2Fe-2S] cluster and subsequently the NAD(H) binding site at S-FAD. Biophysical analysis of the enzyme provides the first direct insight into the mechanism of flavin-based electron bifurcation and the requisite structural features
745843
Pyrococcus furiosus
NADH + H+ + 2 NADP+ + 2 reduced ferredoxin [iron-sulfur] cluster
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
heterodimer
1 * 31000 + 1 * 53000, SDS-PAGE
Pyrococcus furiosus
Other publictions for EC 1.6.1.4
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
745371
Nguyen
Two functionally distinct NAD ...
Pyrococcus furiosus
J. Biol. Chem.
292
14603-14616
2017
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1
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1
1
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745843
Lubner
Mechanistic insights into ene ...
Pyrococcus furiosus
Nat. Chem. Biol.
13
655-659
2017
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1
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1
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1
1
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741703
Tian
Ferredoxin NAD+ oxidoreductas ...
Thermoanaerobacterium saccharolyticum, Thermoanaerobacterium saccharolyticum LL1025
Appl. Environ. Microbiol.
82
7134-7141
2016
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742880
Demmer
Insights into flavin-based el ...
Thermotoga maritima, Thermotoga maritima ATCC 43589
J. Biol. Chem.
290
21985-21995
2015
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1
1
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1
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2
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742779
Lewis
Role for ferredoxin NAD(P)H o ...
Pseudomonas putida, Pseudomonas putida DSM 3601
J. Bacteriol.
195
3876-3887
2013
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734518
Cho
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Improvement of hydrogen produc ...
Escherichia coli
J. Nanoelectr. Optoelectr.
6
343-347
2011
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1
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731998
Wang
NADP+ reduction with reduced f ...
Clostridium kluyveri, Clostridium kluyveri DSM 555
J. Bacteriol.
192
5115-5123
2010
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1
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2
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6
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6
1
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288551
Kurokawa
-
Purification of a flavoprotein ...
Nitrobacter winogradskyi, Nitrobacter winogradskyi ATCC 14123
Arch. Microbiol.
148
95-99
1987
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