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Information on EC 1.18.6.1 - nitrogenase and Organism(s) Azotobacter vinelandii and UniProt Accession P00459
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1.18.6.1 nitrogenaseIUBMB Comments
Requires Mg2+. The enzyme is a complex of two components (namely dinitrogen reductase and dinitrogenase). Dinitrogen reductase is a [4Fe-4S] protein, which, in the presence of two molecules of ATP, transfers an electron from ferredoxin to the dinitrogenase component. Dinitrogenase is a molybdenum-iron protein that reduces dinitrogen to two molecules of ammonia in three successive two-electron reductions via diazene and hydrazine. The reduction is initiated by formation of hydrogen in stoichiometric amounts . Acetylene is reduced to ethylene (but only very slowly to ethane), azide to nitrogen and ammonia, and cyanide to methane and ammonia. In the absence of a suitable substrate, hydrogen is slowly formed. Ferredoxin may be replaced by flavodoxin [see EC 1.19.6.1 nitrogenase (flavodoxin)]. The enzyme does not reduce CO (cf. EC 1.18.6.2, vanadium-dependent nitrogenase).
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Word Map
- 1.18.6.1
- nodule
- azotobacter
-
nitrogen-fixing
- vinelandii
-
diazotrophic
- acetylene
-
symbiotic
- klebsiella
- ammonia
- pneumoniae
- cyanobacterium
- rhizobium
- anabaena
- heterocysts
- nitrate
-
legume
-
n2-fixing
-
bacteroid
- hydrogenase
- epr
-
free-living
-
atmospheric
- nh3
- rubrum
- bradyrhizobium
- azospirillum
- rhodospirillum
- japonicum
- rhodobacter
- meliloti
-
rhizosphere
- nostoc
- capsulatus
- pasteurianum
- brasilense
-
microaerobic
- rhodopseudomonas
- leguminosarum
- leghemoglobin
- variabilis
- mgadp
-
carbide
-
dithionite-reduced
-
endor
-
biofertilizer
- flavodoxins
-
drag
-
symbioses
- bacteriochlorophyll
- synthesis
- agriculture
- energy production
-
anoxygenic
The taxonomic range for the selected organisms is: Azotobacter vinelandii
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea
Reaction Schemes
4
+
8
+
+
16
+
16
=
4
+
+
2
+
16
+
16
Synonyms
nitrogenase, mofe protein, dinitrogenase, nifhdk, nifdk, mo-nitrogenase, mofe-protein, v-nitrogenase, nitrogenase mofe protein, n2ase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
Mo-nitrogenase
MoFe protein
nitrogenase Fe protein
nitrogenase MoFe protein
nitrogenase MoFe-protein
additional information
-
the nitrogenase complex is composed of 2 oxygen-labile metalloproteins: dinitrogenase and dinitrogenase reductase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
overall catalytic mechanism, overview, structures of active site metal clusters, interactions of substrate and active site, active site relevant residues are Arg96, Val70, Gly69, and His195, substrate binding mechanism of complex components, mechanism of MgATP hydrolysis and electron transfer, overview
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
NO2- reduction by nitrogenase begins with the generation of NO2H bound to a state in which the active-site FeMo-cofactor (M) has accumulated two [e-/H+] (E2), stored as a (bridging) hydride and proton. Proton transfer to NO2H and H2O loss leaves M-[NO+], transfer of the E2 hydride to the [NO+] directly to form HNO bound to FeMo-cofactor is one of two alternative means for avoiding formation of a terminal M-[NO] thermodynamic sink. Mechanism for N2 reduction, detailed overview. NO2- reduction by nitrogenase begins with the generation of NO2H bound to E2, E2 has accumulated two [e-/H+], stored in the form of a hydride bridging between two Fe atoms and a proton bound to sulfur. Transfer of the E2 proton to the -OH of NO2H followed by loss of H2O formally leaves M-[NO+]. Nitrogenase is able to transfer the E2 hydride to [NO+], directly forming HNO bound to FeMo-cofactor at its resting-state redox level and totally avoiding formation of an M-[NO] sink
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
catalytic mechanism
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
mechanism, Fe protein cycle
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
enzyme complex dissociation and association kinetics
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
C2H2 inhibition mechanism, structure model
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
MgATP/MgADP-dependent electron and proton transfer kinetics
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
structure of V-containing enzyme form
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
enzyme is composed of 2 metalloproteins: component I MoFe protein and component II Fe protein
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
Fe protein and MoFe protein are assumed to associate and dissociate to transfer a single electron to the substrates
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
schematic electron flow from Fe protein to substrate via MoFe protein and MoFe protein-cofactor
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
schematic mechanism
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
active site is located on the MoFe cofactor involving residues alphaR96, alphaG69, alphaV70, and alphaH195
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
active site location
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
theoretical mechanisms of substrate binding to molybdenum or iron in the FeMo cofactor, modeling
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
first introduction of H at the NFe7MoS9(homocitrate) active site is via a water chain terminating at water 679 to S3B of the my3-S atoms of the active site. Discussion of subsequent movement of the H atoms around the NFe7MoS9(homocitrate) preparatory to the binding and hydrogenation of N2 and other substrates. S2B has a modulatory function and is not an H-entry site
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
SEPR1 intermediate formed during turnover of the nitrogenase alpha-195Gln MoFe protein with C2H2 in H2O buffers, is a product complex with C2H4 bound as a ferracycle to a single Fe of the FeMo-cofactor active site. CO bridges two Fe of lo-Co, while the C2H4 of SEPR1 binds to one of these. Correlation with Lowe-Thorneley En kinetic state
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8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
theoretical investigation of the binding of N2 to the Fe7MoS9N(homocitrate)(cysteine)(histidine) active site, calculation of reaction profiles and activation energies for the association and dissociation of N2. An endo-ny1-N2 coordination at Fe6 is most probable
-
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
reaction mechanism, determination of reaction intermediates, two-step relaxation of the nitrogenase H+/H+ intermediate during step-annealing,both steps show large solvent kinetic isotope effects, step A is the catalytically central state that is activated for N2 binding by the accumulation of 4 electrons, and step B accumulates 2 electrons, overview
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REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
redox reaction
-
-
-
-
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
BRENDA
KEGG
-
-
SYSTEMATIC NAME
IUBMB Comments
ferredoxin:dinitrogen oxidoreductase (ATP-hydrolysing, molybdenum-dependent)
Requires Mg2+. The enzyme is a complex of two components (namely dinitrogen reductase and dinitrogenase). Dinitrogen reductase is a [4Fe-4S] protein, which, in the presence of two molecules of ATP, transfers an electron from ferredoxin to the dinitrogenase component. Dinitrogenase is a molybdenum-iron protein that reduces dinitrogen to two molecules of ammonia in three successive two-electron reductions via diazene and hydrazine. The reduction is initiated by formation of hydrogen in stoichiometric amounts [2]. Acetylene is reduced to ethylene (but only very slowly to ethane), azide to nitrogen and ammonia, and cyanide to methane and ammonia. In the absence of a suitable substrate, hydrogen is slowly formed. Ferredoxin may be replaced by flavodoxin [see EC 1.19.6.1 nitrogenase (flavodoxin)]. The enzyme does not reduce CO (cf. EC 1.18.6.2, vanadium-dependent nitrogenase).
CAS REGISTRY NUMBER
COMMENTARY
9013-04-1
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
overall reaction
-
-
?
hydrazine + reduced ferredoxin
2 NH3 + oxidized ferredoxin
-
-
-
?
hydroxylamine + reduced ferredoxin
NH3 + H2O + oxidized ferredoxin
-
-
-
?
N2 + 4 reduced ferredoxin
hydrazine + 4 oxidized ferredoxin
-
-
-
?
nitrite + 4 reduced ferredoxin + 5 H+
hydroxylamine + H2O + 4 oxidized ferredoxin
-
-
-
?
nitrite + 7 H+ + 12 ATP
NH3 + 2 H2O + 12 ADP + 12 phosphate
overall reaction
-
-
?
nitrite + H+ + ATP + reduced ferredoxin
NH3 + 2 H2O + 12 ADP + 12 phosphate + oxidized ferredoxin
overall reaction
-
-
?
(VO4)3- + ?
(VO2)+ + ?
-
reduction of vanadium(V) by reduced Fe-protein of enzyme to vanadium(IV), which then probably binds to the nucleotide binding site in place of the Mg2+ which is normally present. The oxidized Fe-protein is unable to reduce vanadate
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
3,3-difluorocyclopropene + dithionite
propene + 2-fluoropropene + ?
-
-
major products providing evidence for reductive C-F bond cleavage. Synthesis of propene requires 6 e-/6 H+ and of 2-fluoropropene requires 4 e-/4 H+. In both products, C=C bond cleavage rather than C-C bond cleavage is involved. No selectivity is observed in formation of cis and trans isomers of 1,3-d2-2-fluoropropene, whereas cis-1,3-d2-propene is the predominant 1,3-d2-propene product, indicating that one of the bound reduction intermediates on the pathway to propene is constrained geometrically. Reduction requires both N2ase proteins MoFe and Fe protein, ATP, and an exogenous reductant such as dithionite. A reduction mechanism, consistent with hydride transfer as a key step, is discussed
-
?
6 reduced flavodoxin + N2 + 6 H2O + 6 ATP
6 oxidized flavodoxin + 2 NH3 + 6 H+ + 6 ADP + 6 phosphate
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
CN- + europium(II) diethylenetriaminepentaacetate + H+
?
-
-
cofactor-deficient enzyme catalyzes formation of methane, ethane, ethene, propane, propene, butene, butane, pentene, pentane
-
?
CO + dithionite + H+
CH4 + C2H6 + C2H4 + C3H6 + C3H8 + ?
-
no substrate for wild-type
MoFe subunit mutants V70A and V70G will catalyze the reduction and coupling of CO to form methane, ethane, ethylene, propene, and propane. The rates and ratios of hydrocarbon production from CO can be adjusted by changing the flux of electrons through nitrogenase, by substitution of other amino acids located near the FeMo-cofactor, or by changing the partial pressure of CO. Increasing the partial pressure of CO shifts the product ratio in favor of the longer chain alkanes and alkenes
-
?
CO + europium(II) diethylenetriaminepentaacetate + H+
?
-
-
cofactor-deficient enzyme catalyzes formation of methane, ethane, ethene, propane, propene
-
?
diazene + ?
?
-
modified nitrogenase variant V70A/H195Q under turnover conditions using diazene, methyldiazene, or hydrazine as substrate traps a common S = 1/2 intermediate. Such samples also contain a common intermediate with FeMo-co in an integer-spin state having a ground-state non-Kramers doublet. This species, designated H, has NH2 bound to FeMo-co and corresponds to the penultimate intermediate of N2 hydrogenation, the state formed after the accumulation of seven electrons/protons and the release of the first NH3, the S = 1/2 species corresponds to the final intermediate in N2 reduction, the state formed after accumulation of eight electrons/protons, with NH3 still bound to FeMo-co prior to release and regeneration of resting-state FeMo-co
-
-
?
dithionite + H+ + N2 + ATP + CO2
CO + CH4 + C2H6 + C3H6 + C3H8 + C4H8 + C4H10 + ?
-
-
-
-
?
N2 + 10 H+ + 8 e- + 16 ATP
2 NH4+ + H2 + 16 ADP + 16 phosphate
-
-
-
-
?
propargyl alcohol + ?
?
-
wild-type enzyme and V70A mutant MoFe protein-containing enzyme
-
-
?
reduced ferredoxin + H+ + CH3NC + ATP
oxidized ferredoxin + CH4 + C2H4 + C3H6 + C3H8 + CH3NH2 + ADP + phosphate
reduced ferredoxin + H+ + N2O + ATP
oxidized ferredoxin + H2O + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + SCN- + ATP
oxidized ferredoxin + H2S + HCN + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
enzyme complex is responsible for the majority of biological nitrogen fixation
-
-
ir
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
turnover cycle scheme, MgATP is required for activity, mechanism of MgATP hydrolysis and electron transfer with an important role of switch I and II within the Fe protein
-
-
ir
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
active site for acetylene reduction interacts not directly with N2 reduction
-
-
?
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
anaerobic atmosphere
reduction cycle continues until complete reduction of the substrate to ethane
?
6 reduced flavodoxin + N2 + 6 H2O + 6 ATP
6 oxidized flavodoxin + 2 NH3 + 6 H+ + 6 ADP + 6 phosphate
-
-
-
-
?
6 reduced flavodoxin + N2 + 6 H2O + 6 ATP
6 oxidized flavodoxin + 2 NH3 + 6 H+ + 6 ADP + 6 phosphate
-
intermediate is a flavodoxin hydroquinone
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
slow enzyme
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
1-propyne, 1-butyne and allene are reduced to the corresponding alkenes
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
Fe protein and MoFe protein are assumed to associate and dissociate to transfer a single electron to the substrates, termed Fe protein cycle, driven by MgATP hydrolysis, with the dissociation of the Fe protein-MoFe protein complex being the rate limiting step of the cycle
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
MgATP-dependent
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
MgATP-dependent
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
MgATP-dependent
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
biological N2 fixation
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
biological N2 fixation
-
?
dithionite + H+ + N2 + ATP
?
-
in vitro substrate
-
-
?
dithionite + H+ + N2 + ATP
?
-
SO2- being the actual nitrogenase reductant, reaction kinetics
-
-
?
hydrazine + ?
?
-
high activity with the Val70 mutant enzyme, poor substrate for the wild-type enzyme
-
-
?
N2 + 8 e- + 16 ATP + 8 H+
2 NH3 + H2 + 16 ADP + 16 phosphate
-
-
-
-
?
N2 + 8 e- + 16 ATP + 8 H+
2 NH3 + H2 + 16 ADP + 16 phosphate
-
the enzyme is responsible for biological nitrogen fixation, the conversion of atmospheric N2 to NH3
-
-
?
N2 + 8 e- + 16 ATP + 8 H+
2 NH3 + H2 + 16 ADP + 16 phosphate
-
relaxation of the nitrogenase H+/H+ intermediate during step-annealing
-
-
?
N2 + 8 e- + 8 H+ + 16 ATP
2 NH3 + H2 + 16 ADP + 16 phosphate
-
-
-
-
ir
N2 + 8 e- + 8 H+ + 16 ATP
2 NH3 + H2 + 16 ADP + 16 phosphate
-
cofactor binding structure analysis, Fe protein-MoFe protein complex structure in the presence of ATP analogue AMPPCP, overview
-
-
ir
reduced ferredoxin + H+ + ATP
oxidized ferredoxin + H2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + ATP
oxidized ferredoxin + H2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + ATP
oxidized ferredoxin + H2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + ATP
oxidized ferredoxin + H2 + ADP + phosphate
-
in absence of other acceptors
-
-
?
reduced ferredoxin + H+ + CH3NC + ATP
oxidized ferredoxin + CH4 + C2H4 + C3H6 + C3H8 + CH3NH2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CH3NC + ATP
oxidized ferredoxin + CH4 + C2H4 + C3H6 + C3H8 + CH3NH2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CH3NC + ATP
oxidized ferredoxin + CH4 + C2H4 + C3H6 + C3H8 + CH3NH2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CH3NC + ATP
oxidized ferredoxin + CH4 + C2H4 + C3H6 + C3H8 + CH3NH2 + ADP + phosphate
-
-
-
-
?
reduced ferredoxin + H+ + CN- + ATP
oxidized ferredoxin + CH4 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CN- + ATP
oxidized ferredoxin + CH4 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CN- + ATP
oxidized ferredoxin + CH4 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CN- + ATP
oxidized ferredoxin + CH4 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + CN- + ATP
oxidized ferredoxin + CH4 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
-
ir
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
biological nitrogen fixation
-
-
ir
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
in absence of N2 or other substrates, the electron flow is directed towards proton reduction
-
-
ir
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N3- + ATP
oxidized ferredoxin + NH3 + N2 + ADP + phosphate
-
mutant H195Q shows only about 7.5% activity compared to wild-type
-
?
additional information
?
-
the activity of the enzyme complex is regulated by specific interactions, inducing conformational changes, between the complex components, overview
-
-
?
additional information
?
-
catalytic role of the molybdenum-iron protein, with P-cluster, and the Fe protein
-
-
?
additional information
?
-
nitrite and hydroxylamine are nitrogenase substrates. The proposed NO2- reduction intermediate hydroxylamine (NH2OH) is a nitrogenase substrate, reduction intermediates can be trapped, cf. EC 1.7.2.2
-
-
?
additional information
?
-
nitrite and hydroxylamine as nitrogenase substrates. The proposed NO2- reduction intermediate hydroxylamine (NH2OH) is a nitrogenase substrate, reduction intermediates can be trapped, cf. EC 1.7.2.2
-
-
?
additional information
?
-
-
nitrogenase catalyzes the biological reduction of N2 to ammonia as well as the two-electron reduction of the nonphysiological alkyne substrate, alkyne substrate interaction within the nitrogenase MoFe protein, overview, the addition of neither 2-butyne-1-ol nor 2-butyne-1,4-diol to the growth medium has any effect on the capacity of wild-type Azotobacter vinelandii to sustain diazotrophic growth
-
-
?
additional information
?
-
-
substrate specificity of wild-type and mutant enzymes, reduction reactions using acetylene, propyne, 1-butyne, 2-butyne, propargyl alcohol, 2-butyne-1-ol, and 2-butyne-1,4-diol as substrates, overview
-
-
?
additional information
?
-
-
nitrogenase catalyzes the nucleotide-dependent conversion of dinitrogen to ammonia at the iron-molybdenum cofactor center of its molybdenum-iron protein component. Mo and homocitrate can be loaded onto the Fe protein upon ATP hydrolysis. Mo may enter the Fe protein by attaching to the position that corresponds to the gamma-phosphate of ATP following the hydrolysis of ATP. Subsequently, the loaded Fe protein can deliver Mo and homocitrate to the NifEN-associated precursor and transform the precursor into a fully matured iron-molybdenum cofactor
-
-
?
additional information
?
-
substrates bind and are reduced at a single 4Fe-4S face of the FeMo-cofactor. When alpha70Val is substituted by alpha70Ile, access of substrates to Fe6 of this face is effectively blocked
-
-
?
additional information
?
-
-
in the draft mechanism model, H2 is produced by reductive elimination of the two bridging hydrides of a four-electron reduced intermediate during N2 binding. This process releases H2, yielding N2 bound to FeMo-cofactor that is doubly reduced relative to the resting redox level, and thereby is activated to promptly generate bound diazene. This mechanism predicts that during turnover under D2/N2, the reverse reaction of D2 with the N2-bound product of reductive elimination would generate a dideutero-four-electron reduced intermediate, which can relax with loss of HD to the state designated two-electron reduced intermediate, with a single deuteride bridge. The predicted two-electron reduced intermediate(D) and four-electron reduced intermediate(2D) states are established by intercepting them with the nonphysiological substrate acetylene to generate deuterated ethylenes. That gaseous H2/D2 can reduce a substrate other than H+ with N2 as a cocatalyst confirms the essential mechanistic role for H2 formation, and hence a limiting stoichiometry for biological nitrogen fixation of eight electrons/protons
-
-
?
additional information
?
-
-
a variant enzym containing P-clusters which consist of paired [Fe4S4]-like clusters, can catalyze ATP-independent substrate reduction in the presence of a strong reductant, europium (II) diethylenetriaminepentaacetate [Eu(II)-DTPA]. The variant P*-cluster is not only capable of catalyzing the two-electron reduction of proton, acetylene, ethylene, and hydrazine, but also capable of reducing cyanide, carbon monoxide, and carbon dioxide to alkanes and alkenes
-
-
?
additional information
?
-
-
data support a deficit-spending model of electron transfer where the first event is electron tranfer from the P-cluster to FeMo-cofactor and the second, backfill, event is fast electron tranfer from the Fe protein [4Fe-4S] cluster to the oxidized P-cluster. The first electron transfer is conformationally gated, whereas the second is not
-
-
?
additional information
?
-
-
draft mechanism for N2 reduction by nitrogenase. Diazene binds to the one-electron reduced intermediate with the release of H2, and enters the N2 pathway as the final form of the four-electron reduced state. In contrast, N2H4 instead binds to one-electron reduced intermediate, as is proposed for another two-electron substrate, C2H2, and joins the N2 pathway at a stage corresponding to the seven-electron reduced intermediate in the N2 reduction scheme
-
-
?
additional information
?
-
-
using buffer 3-(N-morpholino)propanesulfonic acid, which has a very small temperature coefficient, temperature-dependent elctron transfer rate constants are observed, with nonlinear Arrhenius plots and with electron transfers gated across the temperature range by a conformational change that involves the binding of numerous water molecules, consistent with an unchanging electron transfer mechanism. There is no solvent kinetic isotope effect throughout the temperature range studied, consistent with an unchanging mechanismn. The nonlinear Arrhenius plots are explained by the change in heat capacity caused by the binding of water molecules in an invariant gating electron transfer mechanism. The observations contradict the idea of a change in electron transfer mechanism with cooling
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
overall reaction
-
-
?
hydrazine + reduced ferredoxin
2 NH3 + oxidized ferredoxin
-
-
-
?
hydroxylamine + reduced ferredoxin
NH3 + H2O + oxidized ferredoxin
-
-
-
?
N2 + 4 reduced ferredoxin
hydrazine + 4 oxidized ferredoxin
-
-
-
?
nitrite + 4 reduced ferredoxin + 5 H+
hydroxylamine + H2O + 4 oxidized ferredoxin
-
-
-
?
nitrite + H+ + ATP + reduced ferredoxin
NH3 + 2 H2O + 12 ADP + 12 phosphate + oxidized ferredoxin
overall reaction
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
enzyme complex is responsible for the majority of biological nitrogen fixation
-
-
ir
2 reduced ferredoxin + 2 H+ + acetylene + 2 ATP + 2 H2O
2 oxidized ferredoxin + ethylene + 2 ADP + 2 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
N2 + 10 H+ + 8 e- + 16 ATP
2 NH4+ + H2 + 16 ADP + 16 phosphate
-
-
-
-
?
N2 + 8 e- + 16 ATP + 8 H+
2 NH3 + H2 + 16 ADP + 16 phosphate
-
the enzyme is responsible for biological nitrogen fixation, the conversion of atmospheric N2 to NH3
-
-
?
N2 + 8 e- + 8 H+ + 16 ATP
2 NH3 + H2 + 16 ADP + 16 phosphate
-
-
-
-
ir
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
440138, 440142, 440143, 440144, 440145, 440146, 440147, 440150, 440153, 440154, 440155, 440156, 440158, 440163, 440166
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
-
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
biological N2 fixation
-
-
?
8 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O
8 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate
-
biological N2 fixation
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
-
ir
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
-
-
?
reduced ferredoxin + H+ + N2 + ATP + H2O
oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
biological nitrogen fixation
-
-
ir
additional information
?
-
the activity of the enzyme complex is regulated by specific interactions, inducing conformational changes, between the complex components, overview
-
-
?
additional information
?
-
nitrite and hydroxylamine are nitrogenase substrates. The proposed NO2- reduction intermediate hydroxylamine (NH2OH) is a nitrogenase substrate, reduction intermediates can be trapped, cf. EC 1.7.2.2
-
-
?
additional information
?
-
-
nitrogenase catalyzes the biological reduction of N2 to ammonia as well as the two-electron reduction of the nonphysiological alkyne substrate, alkyne substrate interaction within the nitrogenase MoFe protein, overview, the addition of neither 2-butyne-1-ol nor 2-butyne-1,4-diol to the growth medium has any effect on the capacity of wild-type Azotobacter vinelandii to sustain diazotrophic growth
-
-
?
additional information
?
-
-
nitrogenase catalyzes the nucleotide-dependent conversion of dinitrogen to ammonia at the iron-molybdenum cofactor center of its molybdenum-iron protein component. Mo and homocitrate can be loaded onto the Fe protein upon ATP hydrolysis. Mo may enter the Fe protein by attaching to the position that corresponds to the gamma-phosphate of ATP following the hydrolysis of ATP. Subsequently, the loaded Fe protein can deliver Mo and homocitrate to the NifEN-associated precursor and transform the precursor into a fully matured iron-molybdenum cofactor
-
-
?
additional information
?
-
-
in the draft mechanism model, H2 is produced by reductive elimination of the two bridging hydrides of a four-electron reduced intermediate during N2 binding. This process releases H2, yielding N2 bound to FeMo-cofactor that is doubly reduced relative to the resting redox level, and thereby is activated to promptly generate bound diazene. This mechanism predicts that during turnover under D2/N2, the reverse reaction of D2 with the N2-bound product of reductive elimination would generate a dideutero-four-electron reduced intermediate, which can relax with loss of HD to the state designated two-electron reduced intermediate, with a single deuteride bridge. The predicted two-electron reduced intermediate(D) and four-electron reduced intermediate(2D) states are established by intercepting them with the nonphysiological substrate acetylene to generate deuterated ethylenes. That gaseous H2/D2 can reduce a substrate other than H+ with N2 as a cocatalyst confirms the essential mechanistic role for H2 formation, and hence a limiting stoichiometry for biological nitrogen fixation of eight electrons/protons
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
FeMo cofactor
active site located, ability of the multimetallic catalytic FeMo cofactor cluster to accumulate multiple [e-/H+]
[4Fe-4S]-center
-
NifB and NifEN are two essential elements immediately adjacent to each other along the biosynthetic pathway of FeMoco. The 8Fe-precursor is present in the NifEN entity of a synthetic NifEN-B fusion protein, and additional [Fe4S4]-type cluster species are present in the NifB entity of NifEN-B. The cluster species in NifEN-B consist of both SAM-motif- and non-SAM-motif-bound [Fe4S4]-type clusters. The non-SAM-motif [Fe4S4]-cluster is a NifB-bound intermediate of FeMoco assembly, which could be converted to the 8Fe-precursor in a SAM-dependent mechanism
ATP
2 MgATP binding sites on the iron protein, binding changes the redox status of the [4Fe4S] cluster of the iron protein, mechanism, overview
FeMo cofactor
-
structure, overview, a complex metallo-organic species called FeMo-cofactor provides the site of substrate reduction within the MoFe protein
FeMo cofactor
-
NifEN plays an essential role in the biosynthesis of the nitrogenase iron-molybdenum, FeMo, cofactor. It is an alpha2beta2 tetramer that is homologous to the catalytic MoFe protein, NifDK, component of nitrogenase. NifEN serves as a scaffold for the conversion of an iron-only precursor to a matured form of the M cluster before delivering the latter to its target location within NifDK, NifEN crystal structure analysis, overview
FeMo protein
-
a complex metallo-organic species called FeMo-cofactor provides the site of substrate reduction within the MoFe protein, Fe6 within FeMo-cofactor provides the unique site for alkyne substrate binding and has van der Waals contact with the side chains of alpha-Val70l and alpha-Gln191, overview
-
FeMo protein
-
FeMo-co is composed of 7Fe, 9S, Mo, R-homocitrate, and one unidentified light atom, in vitro synthesis of the iron-molybdenum cofactor of nitrogenase from iron, sulfur, molybdenum, and homocitrate using purified Nif proteins, Several nif genes are essential for FeMo-co synthesis in vivo, e.g., nifB, nifU, nifS, nifH, nifN, nifE, and nifV, modeling, overview
-
iron-molybdenum cofactor
-
dependent on, the enzyme complex contains a molybdenum-iron protein harboring the active site, the cofactor is composed of a [Mo-3Fe-3S] subcluster and a [4Fe3S] subcluster bridged by 3 sulfide pairs, with homocitrate bound to the molybdenum, structure determination and analysis
iron-molybdenum cofactor
-
enzyme contains a [7Fe9S-Mo-X-homocitrate] metallocluster, structure, redox status, part of the MoFe protein
iron-molybdenum cofactor
-
required for activity of the enzyme complex, insertion in the presynthesized apodinitrogenase involving the monomeric 26 kDa NafY protein, which binds the FeMo cofactor with very high affinity via its HIs121, the cofactor-NafY-complex exhibits an EPR signal similar to isolated FeMo cofactor and the M-center of the enzyme, NafY also binds the biosynthetic precursor of the MoFe cofactor, the NifB-cofacor, determination of binding sites
iron-molybdenum cofactor
-
substrate interaction at an iron-sulfur face of the FeMo-cofactor during nitrogenase catalysis
iron-molybdenum cofactor
modelling of the FeMo-cofactor binding site in the alpha70Ile MoFe protein structure
iron-molybdenum cofactor
-
molybdenum-iron protein component with an iron-molybdenum cofactor center, biosynthesis of the cofactor, detailed overview. NifS and NifU launch the process by synthesizing small Fe/S fragments, such as the [Fe2S2] clusters and the [Fe4S4] clusters. These small Fe/S building blocks are assembled into a large Fe/S core on NifB and further processed on NifEN with the assistance of Fe protein. Upon the completion of assembly on NifEN, the mature FeMoco is subsequently delivered to its target location within theMoFe protein, resulting in the formation of an active holo-MoFe protein
iron-molybdenum cofactor
the enzyme contains 2 [8Fe-7S] P clusters and 2 active site [7Fe-9S-C-Mo-homocitrate] iron-molybdenum cofactors
additional information
-
structure models of Fe protein, MoFe protein, and MoFe-cofactor, and their metal centers, e.g. the [4Fe-4S] cluster based on crystallographic data, substrate binding and electron transfer
-
additional information
-
NifEN is a partially defective homologue of the MoFe protein, catalytic properties, overview. Components of the electron transfer chains in nitrogenase and its homologue, overview
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Iron
dependent on, the enzyme complex contains a molybdenum-iron, a tetramer with 2 different subunits and 4 organo-metallic clusters, i.e. 2 iron-molybdenum cofactors and 2 P-clusters encoded by the genes nifD and nifK, the enzyme complex contains also a dimeric iron protein, encoded by the nifH gene, with a [4Fe4S] cluster between subunits and 2 MgATP binding sites, mechanism of electron transfer between metal clusters, mechanism of switch I and II, complex formation with the MoFe protein, also between different species, overview
Mg2+
required for ATP binding and MgATP hydrolysis, 2 MgATP binding sites on the iron protein connected via residues K15, D125, and C132, binding changes the conformation and the redox status of the [4Fe4S] cluster of the iron protein, mechanism, overview
Molybdenum
Co2+
Fe
-
molybdenum-iron protein component and an iron protein component with an iron-molybdenum cofactor center. Schematic structure of the alpha2-dimeric Fe protein, which contains a [Fe4S4] cluster at the subunit interface and an MgATP binding site within each subunit, overview
Fe2+
Iron
Mg2+
Mn2+
Mo
-
molybdenum-iron protein component with an iron-molybdenum cofactor center. Schematic structureof the alpha2beta2-tetrameric MoFe protein, which contains a pair of unique clusters in each ab-subunit dimer, the P-cluster ([Fe8S7]) at the alphabeta-subunit interface, and the FeMoco ([MoFe7S9X], where X=C,N, or O) within the alpha-subunit
Molybdenum
Ni2+
Vanadium
additional information
Molybdenum
dependent on, the enzyme complex contains a molybdenum-iron, a tetramer with 2 different subunits and 4 organo-metallic clusters, i.e. 2 iron-molybdenum cofactors [7Fe-Mo-9S-X-homocitrate] and 2 P-clusters [8Fe-7S], mechanism of electron transfer between metal clusters, complex formation with the Fe protein, also between different species, overview
Co2+
-
divalent cation requirement is satisfied by Co2+, is best supported by concentrations of divalent cation one-half the concentration of ATP
Fe2+
-
divalent cation requirement is satisfied by Fe2+, is best supported by concentrations of divalent cation one-half the concentration of ATP
Iron
-
-
Iron
-
enzyme consists of 2 proteins: a molybdenum and iron-containing protein, MoFe protein, component I, dinitrogenase, and an iron containing protein, Fe protein, component II, dinitrogenase reductase, together they form the active nitrogenase complex
Iron
-
the MoFe protein contains 2 molybdenum, about 30 iron and 30 inorganic sulphur atoms, 16 of the 30 Fe atoms are associated with S2- in four cubic [4Fe-4S] clusters, the remaining metal atoms are arranged in two copies of a cofactor called FeMo cofactor, FeMoCo, with a minimum stoichiometry of MoFe6S8-9
Iron
-
characterization of the metal clusters in the nitrogenase molybdenum-iron and vanadium-iron proteins
Iron
-
VFe protein form 1 is an incomplete form that contains only 1 cofactor and 1 [4Fe-4S] cluster with an additional [Fe4-S4]-like cluster
Iron
-
dependent on, the enzyme complex contains a molybdenum-iron protein harboring the active site, the enzyme complex contains also a dimeric iron protein
Iron
-
enzyme contains a [7Fe9S-Mo-X-homocitrate] metallocluster, 1 of 2 different models proposes one or more Fe atoms in the Mo cofactor to be responsible for substrate binding
Iron
-
freeze-trapping the FeMo-cofactor in a S=1/2 state with hydrazine as substrate in mutant V70A/H195Q. The trapped intermediate incorporates a hydrazine-derived species bound to the FeMo-cofactor
Iron
contains iron and a Fe-Mo cofactor (the active site, a [7Fe:9S:Mo:C:R-homocitrate] cluster)
Mg2+
-
divalent metal requirement is satisfied by Mg2+, reaction is best supported by concentration of divalent cation one-half the concentration of ATP
Mg2+
-
Mg2+ required for MgATP complex
Mn2+
-
divalent cation requirement is satisfied by Mn2+, is best supported by concentrations of divalent cation one-half the concentration of ATP
Molybdenum
-
-
Molybdenum
-
1-2 gatom per mol of MoFe protein , overview
Molybdenum
-
characterization of the metal clusters in the nitrogenase molybdenum-iron protein
Molybdenum
-
molybdenum metabolism, cofactor synthesis from nif genes , regulation and structure, overview
Molybdenum
-
also possesses Mo-independent nitrogenases: one vanadium containing nitrogenase and another lacking both molybdenum and vanadium
Molybdenum
-
enzyme consists of 2 proteins: a molybdenum and iron-containing protein (MoFe protein, component I, dinitrogenase) and an iron containing protein (Fe protein, component II, dinitrogenase reductase), together they form the active nitrogenase complex
Molybdenum
-
MoFe-cofactor contains 2 clusters of composition [4Fe-3S] and [1Mo-3Fe-3S] that are brigded by 3 nonprotein ligands
Molybdenum
-
mol Mo per mol MoFeprotein: wild-type and mutant H195Q 1.9, mutant H195N and Q191K 0.9
Molybdenum
-
dependent on, the enzyme complex contains a molybdenum-iron protein harboring the active site, the cofactor is composed of a [Mo-3Fe-3S] subcluster and a [4Fe3S] subcluster bridged by 3 sulfide pairs, with homocitrate bound to the molybdenum, structure determination and analysis
Molybdenum
-
enzyme contains a [7Fe9S-Mo-X-homocitrate] metallocluster, where X can be an N atom, 1 of 2 different models proposes molydenum as the substrate binding partner in the active site
Molybdenum
-
freeze-trapping the FeMo-cofactor in a S=1/2 state with hydrazine as substrate in mutant V70A/H195Q. The trapped intermediate incorporates a hydrazine-derived species bound to the FeMo-cofactor
Molybdenum
-
part of the iron-molybdenum and molybdenum-iron cofactors
Molybdenum
contains a Fe-Mo cofactor (the active site, a [7Fe:9S:Mo:C:R-homocitrate] cluster)
Ni2+
-
divalent cation requirement is satisfied by Ni2+, is best supported by concentrations of divalent cation one-half the concentration of ATP
Vanadium
-
VFe protein form 1 is an incomplete form that contains only 1 cofactor and 1 [4Fe-4S] cluster with an additional [Fe4-S4]-like cluster
Vanadium
-
2 forms of VFe protein: form 1 has V-toFe ratio of 1:19, form 2 of 1:15
Vanadium
-
characterization of the metal clusters in the nitrogenase vanadium-iron protein
Vanadium
-
possesses 2 molybdenum-independent nitrogenases: one vanadium-containing nitrogenase and another lacking both molybdenum and vanadium
additional information
structure catalytic role, and mechanism of the P-cluster, part of the MoFe protein, which has a role in immediate electron acceptance from the Fe protein
additional information
-
other metal ions, e.g. Cu2+, Mg2+, Zn2+, Ca2+, at levels of 1-2 atoms per mol detected in the MoFe protein, no evidence for specific requirement, except for Mg2+ in MgATP complex, of any of these metals
additional information
-
structures of the metal centers in Fe protein and MoFe protein, overview
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Acetylene
noncompetitive inhibition of nitrogen reduction, Gly69 is important
beryllium fluoride
inhibits by trapping of a stable Fe protein-MoFe protein nitrogenase complex
H2
H2 competes with N2 binding and inhibits N2 reduction by the FeMo protein, but H2 does not inhibit NO2- reduction for the wild-type or either of the two MoFe protein variants
tetrafluoroaluminate
inhibits by trapping of a stable Fe protein-MoFe protein nitrogenase complex, binds to the Fe protein
Vanadium
-
contains 3 classes of nitrogenase, the second contains V, the third is encoded by a separate set of genes and is inhibited by V and Mo
CO
-
noncompetitive inhibitor of N2, C2H2 and N3- reduction, no inhibition of H+ reduction
H2
-
competitive inhibitor of N2, no inhibition of N3-, C2H2, CN- or H+ reduction
N2
-
maximal inhibition of H2 production at Fe protein to MoFe protein ration 2.5
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
C2H2
-
enhances the CH4 production but not NH3 production from CN-, wild-type enzyme
homocitrate
-
plays role in electron transfer at the [4Fe-4S] cluster to the MoFe-cofactor of the MoFe protein, can be substituted by erythro-fluorohomocitrate but not by threo-fluorohomocitrate
light
-
light-driven activation of the molybdenum-iron-protein, MoFeP, of nitrogenase for substrate reduction is independent of ATP hydrolysis and the iron-protein, FeP, binding structure and mechanism, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
redox properties of metal clusters and P-cluster, overview
-
additional information
additional information
-
overview, different substrates
-
additional information
additional information
-
determination of EPR signals of wild-type enzyme and V70A mutant MoFe protein-containing enzyme with propargyl alcohol and C2H2 as substrates, temperature dependence
-
additional information
additional information
-
Km for N2 of wild-type and mutant enzyme
-
additional information
additional information
-
kinetic analysis, overview
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
400
flavodoxin hydroquinone
-
before and after reduction of the nitrogenase complex relatively slow reactions take place, which limits the rate of the Fe protein cycle
-
additional information
additional information
-
1200 per min: proton production of the reduced enzyme, MgATP-dependent
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
2.2
about, micromol/min/mg MoFe protein, mutant enzyme, pH not specified in the publication, temperature not specified in the publication
2.316
about, micromol/min/mg MoFe protein, wild-type enzyme, pH not specified in the publication, temperature not specified in the publication
additional information
additional information
-
wild-type and diverse alpha-His195 MoFe protein mutants
additional information
-
rates of ATP hydrolysis by Mo and V nitrogenases are comparable under CO, which reflected a similar flux of electrons through the two nitrogenases
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
using buffer 3-(N-morpholino)propanesulfonic acid, which has a very small temperature coefficient, temperature-dependent elctron transfer rate constants are observed, with nonlinear Arrhenius plots and with electron transfers gated across the temperature range by a conformational change that involves the binding of numerous water molecules, consistent with an unchanging electron transfer mechanism. There is no solvent kinetic isotope effect throughout the temperature range studied, consistent with an unchanging mechanismn. The nonlinear Arrhenius plots are explained by the change in heat capacity caused by the binding of water molecules in an invariant gating electron transfer mechanism. The observations contradict the idea of a change in electron transfer mechanism with cooling
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
additional information
-
not established, whether the nitrogenase exists in vivo in a specific particle or whether the nitrogenase proteins are bound nonspecifically to the membranes of some cells
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
physiological function
the enzyme is involved in the N2 fixation mechanism, that proceeds via two different pathways, overview. N2 and NO2- reduction pathways converge upon reduction of NH2NH2 and NH2OH bound states to form state H with [-NH2] bound to the FeMo cofactor. Final reduction converts reaction intermediates H to I, with NH3 bound to the FeMo cofactor, supporting a N2 fixation mechanism in which liberation of the first NH3 occurs upon delivery of five [e-/H+] to N2, but a total of seven [e-/H+] to FeMo cofactor when obligate H2 evolution is considered, and not earlier in the reduction process
evolution
metabolism
-
NifB and NifEN are two essential elements immediately adjacent to each other along the biosynthetic pathway of FeMoco. The 8Fe-precursor is present in the NifEN entity of a synthetic NifEN-B fusion protein, and additional [Fe4S4]-type cluster species are present in the NifB entity of NifEN-B. The cluster species in NifEN-B consist of both SAM-motif- and non-SAM-motif-bound [Fe4S4]-type clusters. The non-SAM-motif [Fe4S4]-cluster is a NifB-bound intermediate of FeMoco assembly, which could be converted to the 8Fe-precursor in a SAM-dependent mechanism
physiological function
-
the diminished H2 evolution by V nitrogenase originates from the diversion of electrons toward CO reduction, in contrast to the Mo nitrogenase
additional information
evolution
-
NifEN and MoFe protein have evolved from the replication and divergence of a common ancestral gene. NifEN is catalytically active early on in the course of evolution, when the mantle of earth is likely more reduced. Later, NifEN might have gradually evolved into an effective enzyme with a wide range of substrates, i.e. the MoFe protein, while in the meantime adjusting its own role toward synthesizing a catalytically more powerful cofactor, i.e. the iron-molybdenum cofactor
evolution
-
the ability of V nitrogenase to catalyze both CO and N2 reductions suggests a potential link between the evolution of carbon and nitrogen cycles
additional information
comparison of reaction mecanisms of nitrogenase, EC 1.18.6.1, and multiheme cytochrome c nitrite reductase, ccNIR, EC 1.7.2.2, overview
additional information
-
like the nif-encoded molybdenum nitrogenase, the vnf-encoded V nitrogenase is composed of a specific reductant and a catalytic component. Both nitrogenases use a catalytic mechanism that involves ATP-dependent electron transfer from a reductant, the nifH- or vnfH-encoded Fe protein, to the catalytic component, i.e. nifDK-encoded MoFe protein or vnfDGK-encoded VFe protein, and the reduction of N2 at the cofactor site, i.e. FeMoco or FeVco, of the latter
additional information
-
NifEN plays an essential role in the biosynthesis of the nitrogenase iron-molybdenum, FeMo, cofactor. It is an alpha2beta2 tetramer that is homologous to the catalytic MoFe protein, NifDK, component of nitrogenase. NifEN serves as a scaffold for the conversion of an iron-only precursor to a matured form of the M cluster before delivering the latter to its target location within NifDK, NifEN crystal structure analysis, overview
additional information
structure of the alpha70Ile MoFe protein compared to the alpha70Val wild-type MoFe protein, shows a delta-methyl group of alpha70Val that is positioned over Fe6 within the active site FeMo-cofactor
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
46000
-
NafY protein, gel filtration, sedimentation equilibrium centrifugation
60000
additional information
additional information
-
enzyme consists of 2 proteins: a molybdenum and iron-containing protein, MoFe protein, component I, dinitrogenase, and an iron containing protein, Fe protein, component II, dinitrogenase reductase, together they form the active nitrogenase complex
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
tetramer
additional information
dimer
-
Fe protein is a dimer of 2 identical subunits, MW 27500-34600, various methods, overview
dimer
-
2 * 26141-28000, unassociated NafY protein, sequence calculation and sedimentation equilibrium centrifugation
additional information
overall enzyme complex structure with the MoFe protein, containing FeMo-cofactors and P-clusters, and 2 iron proteins, one with a [4Fe4S] cluster and MgATP, overview, components are encoded by the nif genes
additional information
-
enzyme is composed of 2 metalloproteins: Fe protein and MoFe protein which are assumed to associate and dissociate to transfer a single electron to the substrates
additional information
-
VFe protein form 1 has alphabeta2 conformation, VFe protein form 2 has alpha2beta2 conformation
additional information
-
enzyme consists of 2 proteins: a molybdenum and iron-containing protein (MoFe protein, component I, dinitrogenase) and an iron containing protein, Fe protein, component II, dinitrogenase reductase, together they form the active nitrogenase complex
additional information
-
apo-MoFe protein has a alpha2beta2 subunit composition and interacts with Fe protein, can be rebuilt by addition of FeMo-cofactor
additional information
-
alpha-beta-monomer of the FeMo protein consists of the FeMo cofactor FeMo-co with the substrate reduction site and the [4Fe-4S] cluster
additional information
-
the NafY protein monomerizes upon binding to the FeMo cofactor
additional information
-
components of the electron transfer chains in nitrogenase and its homologue, overview
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure analysis of the iron protein, and of the MoFe protein
crystallized in presence of approx. 5% w/v polyethylene glycol 6000 and 0.2-0.4 M MgCl2 under strictly anaerobic conditions, x-ray analysis
-
mutant enzyme R96Q bound to acetylene, capillary batch diffusion method, using 30% (w/v) polyethylene glycol 4000, 0.1 M Tris (pH 8.0), 0.17 M Na2MoO4, and 0.001 M sodium dithionite
nitrogenase containing alpha70Ile mutant MoFe protein, 38 mg/ml protein is diluted in 50 mM Tris buffer, pH 8.0, and 250 mM NaCl, crystallization in 30% PEG 4000, 100 mM Tris, pH 8.0, 170-190 mM sodium molybdate, and 1 mM dithionite, 3-4 weeks, X-ray diffraction structure determination and analysis at 2.3 A resolution, comparison to the wild-type, with alpha70Val crystal structure, PDB ID 1M1N, overview
purified enzyme is diluted at room temperature with 3 volumes of Tris-HCl 0.01 M, pH 7.2, immediate crystal formation
-
sitting drop vapor diffusion method, using 150 mM ACES, 75 mM Tris, 75 mM ethanolamine (pH 9.8), 17-18% (w/v) PEG 3350, 0.8 M NaCl, and 1 mM sodium dithionite
sitting drop vapor diffusion method, using 23% polyethylene glycol 3350, 0.2 M lithium citrate, and 5 mM sodium dithionite
sitting drop vapor diffusion method, using 24-28% (w/v) PEG 8000, 0.75-0.85 M NaCl, 0.1 M imidazole/malate (pH 7.5), 1% glycerol (v/v), 0.5% 2,2,2-trifluoroethanol (v/v) and 2.5 mM Na2S2O4
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D125E
site-directed mutagenesis, mutation alters the properties of the MgATP2- binding site with bound MgADP
G69S
random mutagenesis, beta-subunit residue mutant of the MoFe protein shows highly decreased affinity for acetylene, acetylene inhibits the mutants nitrogen reduction activity in a competitive mode in contrast to the wild-type enzyme
K15Q
site-directed mutagenesis, mutation inhibits the communication of the [4Fe4S] cluster with the MgATP2- binding site
S188C
site-directed mutagenesis, mutation of a residue within the P-cluster of the beta-subunit, alters the EPR signal of the MoFe protein
A175G
-
shows in vivo 55% of enzyme activity compared to wild-type, in vitro 20% activity remaining with purified enzyme, slowlier conformational change upon binding of MgATP, model of steric interactions using x-ray crystal structures
A175S
-
unable to support substrate reduction because of an inability to undergo a required MgATP-induced conformational change
DELTAC153
-
mutation in the MoFe protein of nitrogenase. The rate of oxidation of Fe-protein F1+ to this MoFe protein variant is unchanged from the rate to the wild-type MoFe protein, providing further evidence against a gated hopping electron tansfer model
H195G
-
alpha-His of MoFe protein, site directed mutagenesis, reduced MoFe protein activity, slightly decreased Fe protein activity, altered phenotype
H195L
-
alpha-His of MoFe protein, site directed mutagenesis, reduced MoFe protein activity, increased Fe protein activity, altered phenotype
H195N
H195Q
H195T
-
alpha-His of MoFe protein, site directed mutagenesis, reduced MoFe protein and Fe protein activity, altered phenotype
H195Y
-
alpha-His of MoFe protein, site directed mutagenesis, reduced MoFe protein and Fe protein activity, altered phenotype
Q191A/V70A
-
site-directed mutagenesis, the double mutation does result in significant reduction of 2-butyne, with the exclusive product being 2-cis-butene
Q191K
-
alphaGln191 of MoFe protein, shows 6% activity compared to wild-type, substrate CN-, not affected by addition of C2H2
R96Q
the substitution of Arg to Gln at position 96 makes the active site pocket environment more hydrophobic than that of the native enzyme
S188C
-
mutation in the MoFe protein of nitrogenase. Electron transfer to the MoFe state that contains P-cluster PN and FeMo-cofactor MN is conformationally gated in both wild-type MoFe and S188C mutant MoFe protein and the amino acid substitution S188C does not alter the conformational gate
S69G
-
alpha-subunit MoFe protein, resistant to inhibition by C2H2, thus acetylene binding/reduction site is not directly relevant to the mechanism of nitrogen reduction
V70A
V70A/H195Q
-
mutant used for freeze-trapping the FeMo-cofactor in a S=1/2 state with hydrazine as substrate. The trapped intermediate incorporates a hydrazine-derived species bound to the FeMo-cofactor. EPR and ENDOR analysis of the adduct
V70G
V70I
V70X
-
site-directed mutagenesis, substitution of valine with an amino acid with a smaller side chain increases the hydrazine reduction activity, substitution with an amino acid with a larger side chain decreases the enzyme activity with N2, acetylene or hydrazine
additional information
H195N
-
alphaHis195 of MoFe protein, shows 59% activity compared to wild-type, substrate CN-, NH3 and CH4 production from CN- are decreased by C2H2 addition, NH3 production decreased much less
H195N
-
alpha-His of MoFe protein, site directed mutagenesis, reduced MoFe protein activity, altered phenotype
H195Q
-
below 2% N2 reducing activity remaining compared to wild-type due to less effective N2 binding
H195Q
-
alphaHis195 of MoFe protein, shows 159% activity compared to wild-type, substrate CN-, NH3 and CH4 production from CN- are decreased by C2H2 addition
H195Q
-
alpha-His of MoFe protein, site directed mutagenesis, decreased MoFe protein activity, altered phenotype
H195Q
the mutant shows stronger nuclear resonance vibrational spectroscopy features in the Fe-CO region compared to wild type enzyme
V70A
-
site-directed mutagenesis of an alpha subunit residue of the MoFe cofactor, mutation alters the active site structure, trapping of propargyl alcohol at the active site for structure analysis
V70A
-
site-directed mutagenesis, increased the hydrazine reduction activity, reduced Km comapred to the wild-type enzyme
V70A
-
site-directed mutagenesis, substitution of alpha-70Val by alanine results in an increased capacity for the reduction of the larger alkyne propyne
V70A
-
mutation in MoFe subunit. Mutant protein will catalyze the reduction and coupling of CO to form methane, ethane, ethylene, propene, and propane
V70G
-
site-directed mutagenesis, the mutant MoFe protein variant shows an increased capacity for reduction of the terminal alkyne, 1-butyne, but no detectable reduction of the internal alkyne 2-butyne
V70G
-
mutation in MoFe subunit. Mutant protein will catalyze the reduction and coupling of CO to form methane, ethane, ethylene, propene, and propane
V70I
-
site-directed mutagenesis, decreased the hydrazine reduction activity
V70I
-
site-directed mutagenesis, substitution by isoleucine at this position nearly eliminates the capacity for the reduction of acetylene
V70I
-
the mutant is suitable for analysis of reaction intermediates, since it exhibits the highest concentration of trapped H+-intermediate when turned over under Ar
V70I
substitution of alpha70Val by alpha70Ile results in a MoFe protein that is hampered in its ability to reduce a range of substrates including acetylene and N2, yet retains normal proton reduction activity. The mutant shows H2 evolution of greater than 2200 nmol/min/mg MoFe protein, which is 95% of the wild-type specific activity
additional information
deletion of nifH results in an enzyme complex with a MoFe protein exhibiting altered redox properties and no EPR signal, a Fe protein Lys127 deletion mutant mimics the MgATP-bound-conformation and inhibits nucleotide hydrolyzing activity, formation of nondissociating complex with the MoFe protein
additional information
-
natural nifB deletion mutant, MoFe protein without FeMo-cofactor and with small changes in the electronic properties of the [4Fe-4S] cluster
additional information
-
study of two nifB deletion mutants, having His-tagged MoFe/VFe protein, and two nifH deletion mutants, having His-tagged MoFe proteins, with catalytically active P-cluster variants presumably composed of [4Fe-4S]-like centers that are clearly distinct from the normal P-clusters. Proteins are active in terms of H2 evolution, C2H2 reduction, and N2 fixation upon FeMoco insertion
additional information
-
construction of mutant Azotobacter vinelandii strains DJ1242, DJ1313, and DJ1495, the mutant show loss of the ability to grow under nitrogen fixing conditions, phenotypes, overview
additional information
-
in vitro synthesis of the iron-molybdenum cofactor of nitrogenase using purified proteins, a minimal in vitro system, containing NifB, NifEN, and NifH proteins, together with Fe2+, S2-, MoO4 2-, R-homocitrate, S-adenosyl methionine, and Mg-ATP, is sufficient for the synthesis of FeMo-co and the activation of apo-dinitrogenase under anaerobic-reducing conditions, modeling, overview
additional information
-
a MoFeP variant labeled on its surface with a Ru-photosensitizer is shown to photocatalytically reduce protons and acetylene, most likely at its active site, FeMoco. The uncoupling of nitrogenase catalysis from ATP hydrolysis enables the study of redox dynamics within MoFeP and the population of discrete reaction intermediates, overview
additional information
-
a P-cluster variant, which consists of paired [Fe4S4]-like clusters, can catalyze ATP-independent substrate reduction in the presence of a strong reductant, europium (II) diethylenetriaminepentaacetate [Eu(II)-DTPA]. The observation of a decrease of activity in the rank DELTAnifH, DELTAnifB/DELTAnifZ, and DELTAnifB MoFe protein, corresponds to a decrease of the amount of variant P-clusters in these cofactor-deficient proteins and firmly establishes the variant P-cluster as a catalytically active metal center in Eu(II)-diethylenetriaminepentaacetate-driven reactions. The variant P-cluster is not only capable of catalyzing the two-electron reduction of proton, acetylene, ethylene, and hydrazine, but also capable of reducing cyanide, carbon monoxide, and carbon dioxide to alkanes and alkenes
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
9.5
the enzyme is stable when incubated at pH 9.5. At higher pH values and under turnover conditions, the enzyme is slowly inactivated. Initially the enzyme is reversibly inhibited (about 90%) for substrate reduction at pH 9.5, yet in a second, slower process, the enzyme becomes irreversibly inactivated as measured by substrate reduction at the optimal pH of 7.8. Incubation of the enzyme alone at pH 9.5 shows only minimal activity loss even after 4 h
744320
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
nitrogenase complex is more stable than either the MoFe protein or the Fe protein alone
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
extreme O2 lability, susceptibility to O2 increases with purification, but is retarded in presence of MgCl2
-
440158
overview: O2 lability and protection mechanisms against O2 in various organisms in vivo
-
440166
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant Azotobacter vinelandii NafY protein from Escherichia coli strain BL21(DE3)
-
recombinant wild-type Fe protein, recombinant His-tagged wild-type MoFe protein and recombinant His-tagged MoFe protein mutant V70A, to homogeneity
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of His-tagged wild-type and V70A mutant MoFe protein in strain DJ1310, expression of the wild-type Fe protein
-
overexpression of Azotobacter vinelandii NafY protein in Escherichia coli strain BL21(DE3)
-
the Fe subunit of nitrogenase is expressed in Nicotiana tabacum cultivar Petit Havana
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
energy production
-
the reaction produces H2 as a by-product and is interesting for production of clean energy
synthesis
-
MoFe subunit mutants V70A and V70G will catalyze the reduction and coupling of CO to form methane, ethane, ethylene, propene, and propane. The rates and ratios of hydrocarbon production from CO can be adjusted by changing the flux of electrons through nitrogenase, by substitution of other amino acids located near the FeMo-cofactor, or by changing the partial pressure of CO. Increasing the partial pressure of CO shifts the product ratio in favor of the longer chain alkanes and alkenes
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Haaker, H.; Klugkist, J.
The bioenergetics of electron transport to nitrogenase
FEMS Microbiol. Rev.
46
57-71
1987
Azotobacter vinelandii, Anabaena cylindrica, Anabaena sp., Azospirillum sp., Azotobacter sp., Paenibacillus polymyxa, Chlorobium sp., Chromatium sp., Clostridium pasteurianum, Desulfovibrio sp., Frankia sp., Gloeothece sp., Klebsiella pneumoniae, Leptolyngbya boryana, Rhizobium leguminosarum, Rhizobium sp., Cereibacter sphaeroides, Rhodopseudomonas sp., Rhodospirillum rubrum, Rhizobium sp. ORS 571
-
Duyvis, M.G.; Wassink, H.; Haaker, H.
Nitrogenase of Azotobacter vinelandii: kinetic analysis of the Fe protein redox cycle
Biochemistry
37
17345-17354
1998
Azotobacter vinelandii
Vignais, P.M.; Colbeau, A.; Willison, J.C.; Jouanneau, Y.
Hydrogenase, nitrogenase, and hydrogen metabolism in the photosynthetic bacteria
Adv. Microb. Physiol.
26
155-234
1985
Azotobacter vinelandii, Allochromatium vinosum, Clostridium pasteurianum, Klebsiella pneumoniae, Rhodobacter capsulatus, Rhodospirillum rubrum
Eady, R.R.
Isolation and characterization of various nitrogenases
Methods Enzymol.
69
753-778
1980
Azotobacter vinelandii, Allochromatium vinosum, Azotobacter chroococcum, Paenibacillus polymyxa, Bradyrhizobium japonicum, Clostridium pasteurianum, Klebsiella pneumoniae, Bradyrhizobium lupini, Rhodospirillum rubrum
-
Winter, H.C.; Burris, R.H.
Nitrogenase
Annu. Rev. Biochem.
45
409-426
1976
Azotobacter vinelandii, Allochromatium vinosum, Anabaena cylindrica, Azotobacter chroococcum, Paenibacillus polymyxa, Bradyrhizobium japonicum, Clostridium pasteurianum, Corynebacterium flavescens, Klebsiella pneumoniae, Bradyrhizobium lupini, Rhodospirillum rubrum
Eady, R.R.; Postgate, J.R.
Nitrogenase
Nature
249
805-810
1974
Azotobacter vinelandii, Allochromatium vinosum, Anabaena cylindrica, Azotobacter chroococcum, Paenibacillus polymyxa, Bradyrhizobium japonicum, Clostridium pasteurianum, Corynebacterium flavescens, Desulfovibrio desulfuricans, Escherichia coli, Gloeocapsa sp., Klebsiella pneumoniae, Ornithopus sativus, Leptolyngbya boryana, Bradyrhizobium lupini, Rhizobium sp., Rhodospirillum rubrum, Escherichia coli C-M 74, Corynebacterium flavescens 301
Mortenson, L.E.; Thorneley, R.N.F.
Structure and function of nitrogenase
Annu. Rev. Biochem.
48
387-418
1979
Azotobacter vinelandii, Azotobacter chroococcum, Paenibacillus polymyxa, Clostridium pasteurianum, Klebsiella pneumoniae
Shah, V.K.; Ugalde, R.A.; Imperial, J.; Brill, W.J.
Molybdenum in nitrogenase
Annu. Rev. Biochem.
53
231-257
1984
Azotobacter vinelandii, Paenibacillus polymyxa, Clostridium pasteurianum, Klebsiella pneumoniae
Pau, R.N.
Nitrogenases without molybdenum
Trends Biochem. Sci.
14
183-186
1989
Azotobacter vinelandii, Azotobacter chroococcum
Weininger, M.S.; Mortenson, L.E.
Crystallographic properties of the MoFe proteins of nitrogenase from Clostridium pasteurianum and Azotobacter vinelandii
Proc. Natl. Acad. Sci. USA
79
379-380
1982
Azotobacter vinelandii, Clostridium pasteurianum
-
Burns, R.C.; Hardy, R.W.F.
Purification of nitrogenase and crystallization of its Mo-Fe protein
Methods Enzymol.
24
480-496
1972
Azotobacter vinelandii
Pau, R.N.; Mitchenall, L.A.; Robson, R.L.
Genetic evidence for an Azotobacter vinelandii nitrogenase lacking molybdenum and vanadium
J. Bacteriol.
171
124-129
1989
Azotobacter vinelandii
Morningstar, J.E.; Johnson, M.K.; Case, E.E.; Hales, B.J.
Characterization of the metal clusters in the nitrogenase molybdenum-iron and vanadium-iron proteins of Azotobacter vinelandii using magnetic circular dichroism spectroscopy
Biochemistry
26
1795-1800
1987
Azotobacter vinelandii
Bulen, W.A.; LeComte, J.R.
Nitrogenase complex and its components
Methods Enzymol.
24B
456-470
1972
Azotobacter vinelandii
Burgess, B.K.; Jacobs, D.B.; Stiefel, E.I.
Large-scale purification of high activity Azotobacter vinelandii nitrogenase
Biochim. Biophys. Acta
614
196-209
1980
Azotobacter vinelandii
Oelze, J.
Mechanismen zum Schutz der Sauerstoff-labilen Nitrogenase
Forum Mikrobiol.
4
116-126
1988
Azotobacter vinelandii, Anabaena sp., Trichormus variabilis, Azospirillum brasilense, Azotobacter chroococcum, Azotobacter sp., Frankia sp., Gloeothece sp., Klebsiella pneumoniae, Oscillatoria sp., Rhizobium sp., Rhodospirillum rubrum
-
Zumft, W.G.; Mortenson, L.E.
The nitrogen-fixing complex of bacteria
Biochim. Biophys. Acta
416
1-52
1975
Azotobacter vinelandii, Chromatium sp., Clostridium pasteurianum, Klebsiella pneumoniae, Rhizobium sp.
Duyvis, M.G.; Wassink, H.; Haaker, H.
Pre-steady-state MgATP-dependent proton production and electron transfer by nitrogenase from Azotobacter vinelandii
Eur. J. Biochem.
225
881-890
1994
Azotobacter vinelandii
Rasche, M.E.; Seefeldt, L.C.
Reduction of thiocyanate, cyanate, and carbon disulfide by nitrogenase: kinetic characterization and EPR spectroscopic analysis
Biochemistry
36
8574-8585
1997
Azotobacter vinelandii
Christiansen, J.; Goodwin, P.J.; Lanzilotta, W.N.; Seefeldt, L.C.; Dean, D.R.
Catalytic and biophysical properties of a nitrogenase Apo-MoFe protein produced by a nifB-deletion mutant of Azotobacter vinelandii
Biochemistry
37
12611-12623
1998
Azotobacter vinelandii
Erickson, J.A.; Nyborg, A.C.; Johnson, J.L.; Truscott, S.M.; Gunn, A.; Nordmeyer, F.R.; Watt, G.D.
Enhanced efficiency of ATP hydrolysis during nitrogenase catalysis utilizing reductants That form the all-ferrous redox state of the Fe protein
Biochemistry
38
14279-14285
1999
Azotobacter vinelandii, Clostridium pasteurianum, Azotobacter vinelandii OP
Fisher, K.; Dilworth, M.J.; Kim, C.H.; Newton, W.E.
Azotobacter vinelandii nitrogenases with substitutions in the FeMo-cofactor environment of the MoFe protein: effects of acetylene or ethylene on interactions with H+, HCN, and CN-
Biochemistry
39
10855-10865
2000
Azotobacter vinelandii
Bursey, E.H.; Burgess, B.K.
Characterization of a variant iron protein of nitrogenase that is impaired in its ability to adopt the MgATP-induced conformational change
J. Biol. Chem.
273
16927-16934
1998
Azotobacter vinelandii
Christiansen, J.; Cash, V.L.; Seefeldt, L.C.; Dean, D.R.
Isolation and characterization of an acetylene-resistant nitrogenase
J. Biol. Chem.
275
11459-11464
2000
Azotobacter vinelandii
Dilworth, M.J.; Fisher, K.; Kim, C.H.; Newton, W.E.
Effects on substrate reduction of substitution of histidine-195 by glutamine in the alpha-subunit of the MoFe protein of Azotobacter vinelandii nitrogenase
Biochemistry
37
17495-17505
1998
Azotobacter vinelandii
Kim, J.; Rees, D.C.
Nitrogenase and biological nitrogen fixation
Biochemistry
33
389-397
1994
Azotobacter vinelandii, Azotobacter chroococcum, Clostridium pasteurianum
Johnson, J.L.; Tolley, A.M.; Erickson, J.A.; Watt, G.D.
Steady-state kinetic studies of dithionite utilization, component protein interaction, and the formation of an oxidized iron protein intermediate during Azotobacter vinelandii nitrogenase catalysis
Biochemistry
35
11336-11342
1996
Azotobacter vinelandii
Blanchard, C.Z.; Hales, B.J.
Isolation of two forms of the nitrogenase VFe protein from Azotobacter vinelandii
Biochemistry
35
472-478
1996
Azotobacter vinelandii
Kim, C.H.; Newton, W.E.; Dean, D.R.
Role of the MoFe protein alpha-subunit histidine-195 residue in FeMo-cofactor binding and nitrogenase catalysis
Biochemistry
34
2798-2808
1995
Azotobacter vinelandii
Benton, P.M.; Laryukhin, M.; Mayer, S.M.; Hoffman, B.M.; Dean, D.R.; Seefeldt, L.C.
Localization of a substrate binding site on the FeMo-cofactor in nitrogenase: trapping propargyl alcohol with an alpha-70-substituted MoFe protein
Biochemistry
42
9102-9109
2003
Azotobacter vinelandii, Azotobacter vinelandii DJ1310
Seefeldt, L.C.; Dance, I.G.; Dean, D.R.
Substrate interactions with nitrogenase: Fe versus Mo
Biochemistry
43
1401-1409
2004
Azotobacter vinelandii
Igarashi, R.Y.; Seefeldt, L.C.
Nitrogen fixation: the mechanism of the Mo-dependent nitrogenase
Crit. Rev. Biochem. Mol. Biol.
38
351-384
2003
Azotobacter vinelandii (P00459)
Rubio, L.M.; Singer, S.W.; Ludden, P.W.
Purification and characterization of NafY (apodinitrogenase gamma subunit) from Azotobacter vinelandii
J. Biol. Chem.
279
19739-19746
2004
Azotobacter vinelandii, Azotobacter vinelandii DJ
Barney, B.M.; Igarashi, R.Y.; Dos Santos, P.C.; Dean, D.R.; Seefeldt, L.C.
Substrate interaction at an iron-sulfur face of the FeMo-cofactor during nitrogenase catalysis
J. Biol. Chem.
279
53621-53624
2004
Azotobacter vinelandii
Barney, B.M.; Laryukhin, M.; Igarashi, R.Y.; Lee, H.I.; Dos Santos, P.C.; Yang, T.C.; Hoffman, B.M.; Dean, D.R.; Seefeldt, L.C.
Trapping a hydrazine reduction intermediate on the nitrogenase active site
Biochemistry
44
8030-8037
2005
Azotobacter vinelandii
Fisher, K.; Lowe, D.J.; Petersen, J.
Vanadium (V) is reduced by the 'as isolated' nitrogenase Fe-protein at neutral pH
Chem. Commun. (Camb. )
2006
2807-2809
2006
Azotobacter vinelandii
Dance, I.
The hydrogen chemistry of the FeMo-co active site of nitrogenase
J. Am. Chem. Soc.
127
10925-10942
2005
Azotobacter vinelandii
Lee, H.I.; Sorlie, M.; Christiansen, J.; Yang, T.C.; Shao, J.; Dean, D.R.; Hales, B.J.; Hoffman, B.M.
Electron inventory, kinetic assignment (E(n)), structure, and bonding of nitrogenase turnover intermediates with C2H2 and CO
J. Am. Chem. Soc.
127
15880-15890
2005
Azotobacter vinelandii
Dance, I.
The mechanistically significant coordination chemistry of dinitrogen at FeMo-co, the catalytic site of nitrogenase
J. Am. Chem. Soc.
129
1076-1088
2007
Azotobacter vinelandii
Hu, Y.; Corbett, M.C.; Fay, A.W.; Webber, J.A.; Hedman, B.; Hodgson, K.O.; Ribbe, M.W.
Nitrogenase reactivity with P-cluster variants
Proc. Natl. Acad. Sci. USA
102
13825-13830
2005
Azotobacter vinelandii
Dos Santos, P.C.; Mayer, S.M.; Barney, B.M.; Seefeldt, L.C.; Dean, D.R.
Alkyne substrate interaction within the nitrogenase MoFe protein
J. Inorg. Biochem.
101
1642-1648
2007
Azotobacter vinelandii, Azotobacter vinelandii DJ995
Curatti, L.; Hernandez, J.A.; Igarashi, R.Y.; Soboh, B.; Zhao, D.; Rubio, L.M.
In vitro synthesis of the iron-molybdenum cofactor of nitrogenase from iron, sulfur, molybdenum, and homocitrate using purified proteins
Proc. Natl. Acad. Sci. USA
104
17626-17631
2007
Azotobacter vinelandii
Hu, Y.; Fay, A.W.; Lee, C.C.; Wiig, J.A.; Ribbe, M.W.
Dual functions of NifEN: insights into the evolution and mechanism of nitrogenase
Dalton Trans.
39
2964-2971
2010
Azotobacter vinelandii
Roth, L.E.; Nguyen, J.C.; Tezcan, F.A.
ATP- and iron-protein-independent activation of nitrogenase catalysis by light
J. Am. Chem. Soc.
132
13672-13674
2010
Azotobacter vinelandii
Sarma, R.; Barney, B.; Keable, S.; Dean, D.; Seefeldt, L.; Peters, J.
Insights into substrate binding at FeMo-cofactor in nitrogenase from the structure of an alpha70Ile MoFe protein variant
J. Inorg. Biochem.
104
385-389
2010
Azotobacter vinelandii (P07328)
Lee, C.C.; Hu, Y.; Ribbe, M.W.
Vanadium nitrogenase reduces CO
Science
329
642
2010
Azotobacter vinelandii
Kaiser, J.T.; Hu, Y.; Wiig, J.A.; Rees, D.C.; Ribbe, M.W.
Structure of precursor-bound NifEN: a nitrogenase FeMo cofactor maturase/insertase
Science
331
91-94
2011
Azotobacter vinelandii
Danyal, K.; Dean, D.R.; Hoffman, B.M.; Seefeldt, L.C.
Electron transfer within nitrogenase: evidence for a deficit-spending mechanism
Biochemistry
50
9255-9263
2011
Azotobacter vinelandii, Azotobacter vinelandii DJ995
Mayweather, D.; Danyal, K.; Dean, D.R.; Seefeldt, L.C.; Hoffman, B.M.
Temperature invariance of the nitrogenase electron transfer mechanism
Biochemistry
51
8391-8398
2012
Azotobacter vinelandii, Azotobacter vinelandii DJ995
Ni, F.; Lee, C.C.; Hwang, C.S.; Hu, Y.; Ribbe, M.W.; McKenna, C.E.
Reduction of fluorinated cyclopropene by nitrogenase
J. Am. Chem. Soc.
135
10346-10352
2013
Azotobacter vinelandii, Azotobacter vinelandii OP
Yang, Z.Y.; Dean, D.R.; Seefeldt, L.C.
Molybdenum nitrogenase catalyzes the reduction and coupling of CO to form hydrocarbons
J. Biol. Chem.
286
19417-19421
2011
Azotobacter vinelandii, Azotobacter vinelandii DJ1260
Wiig, J.A.; Hu, Y.; Ribbe, M.W.
NifEN-B complex of Azotobacter vinelandii is fully functional in nitrogenase FeMo cofactor assembly
Proc. Natl. Acad. Sci. USA
108
8623-8627
2011
Azotobacter vinelandii
Lukoyanov, D.; Yang, Z.Y.; Barney, B.M.; Dean, D.R.; Seefeldt, L.C.; Hoffman, B.M.
Unification of reaction pathway and kinetic scheme for N2 reduction catalyzed by nitrogenase
Proc. Natl. Acad. Sci. USA
109
5583-5587
2012
Azotobacter vinelandii, Azotobacter vinelandii DJ1260
Lee, C.C.; Hu, Y.; Ribbe, M.W.
ATP-independent substrate reduction by nitrogenase P-cluster variant
Proc. Natl. Acad. Sci. USA
109
6922-6926
2012
Azotobacter vinelandii, Azotobacter vinelandii DJ1143
Yang, Z.Y.; Khadka, N.; Lukoyanov, D.; Hoffman, B.M.; Dean, D.R.; Seefeldt, L.C.
On reversible H2 loss upon N2 binding to FeMo-cofactor of nitrogenase
Proc. Natl. Acad. Sci. USA
110
16327-16332
2013
Azotobacter vinelandii, Azotobacter vinelandii DJ1260
Shaw, S.; Lukoyanov, D.; Danyal, K.; Dean, D.R.; Hoffman, B.M.; Seefeldt, L.C.
Nitrite and hydroxylamine as nitrogenase substrates mechanistic implications for the pathway of N2 reduction
J. Am. Chem. Soc.
136
12776-12783
2014
Azotobacter vinelandii (P00459)
Yang, K.Y.; Haynes, C.A.; Spatzal, T.; Rees, D.C.; Howard, J.B.
Turnover-dependent inactivation of the nitrogenase MoFe-protein at high pH
Biochemistry
53
333-343
2014
Azotobacter vinelandii (P07328)
Morrison, C.N.; Hoy, J.A.; Zhang, L.; Einsle, O.; Rees, D.C.
Substrate pathways in the nitrogenase MoFe protein by experimental identification of small molecule binding sites
Biochemistry
54
2052-2060
2015
Klebsiella pneumoniae (P00466 and P09772), Klebsiella pneumoniae, Clostridium pasteurianum (P00467 and P11347), Clostridium pasteurianum, Azotobacter vinelandii (P07328 and P07329), Azotobacter vinelandii
Sickerman, N.S.; Hu, Y.; Ribbe, M.W.
Activation of CO2 by vanadium nitrogenase
Chem. Asian J.
12
1985-1996
2017
Azotobacter vinelandii
Spatzal, T.; Perez, K.A.; Howard, J.B.; Rees, D.C.
Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor
eLife
4
e11620
2015
Azotobacter vinelandii
Buren, S.; Rubio, L.M.
State of the art in eukaryotic nitrogenase engineering
FEMS Microbiol. Lett.
365
2018
2018
Azotobacter vinelandii
Scott, A.D.; Pelmenschikov, V.; Guo, Y.; Yan, L.; Wang, H.; George, S.J.; Dapper, C.H.; Newton, W.E.; Yoda, Y.; Tanaka, Y.; Cramer, S.P.
Structural characterization of CO-inhibited Mo-nitrogenase by combined application of nuclear resonance vibrational spectroscopy, extended X-ray absorption fine structure, and density functional theory new insights into the effects of CO binding and the
J. Am. Chem. Soc.
136
15942-15954
2014
Azotobacter vinelandii (P07328 and P07329), Azotobacter vinelandii
Lukoyanov, D.; Yang, Z.Y.; Khadka, N.; Dean, D.R.; Seefeldt, L.C.; Hoffman, B.M.
Identification of a key catalytic intermediate demonstrates that nitrogenase is activated by the reversible exchange of N2 for H2
J. Am. Chem. Soc.
137
3610-3615
2015
Azotobacter vinelandii, Azotobacter vinelandii DJ995
Morrison, C.N.; Spatzal, T.; Rees, D.C.
Reversible protonated resting state of the nitrogenase active site
J. Am. Chem. Soc.
139
10856-10862
2017
Azotobacter vinelandii, Clostridium pasteurianum
Hu, Y.; Ribbe, M.W.
A journey into the active center of nitrogenase
J. Biol. Inorg. Chem.
19
731-736
2014
Azotobacter vinelandii, Clostridium pasteurianum
Dance, I.
New insights into the reaction capabilities of His195 adjacent to the active site of nitrogenase
J. Inorg. Biochem.
169
32-43
2017
Azotobacter vinelandii (P07328 and P07329)
Keable, S.; Vertemara, J.; Zadvornyy, O.; Eilers, B.; Danyal, K.; Rasmussen, A.; De Gioia, L.; Zampella, G.; Seefeldt, L.; Peters, J.
Structural characterization of the nitrogenase molybdenum-iron protein with the substrate acetylene trapped near the active site
J. Inorg. Biochem.
180
129-134
2018
Azotobacter vinelandii (P07328 and P07329), Azotobacter vinelandii DJ1264 (P07328 and P07329)
Rebelein, J.G.; Stiebritz, M.T.; Lee, C.C.; Hu, Y.
Activation and reduction of carbon dioxide by nitrogenase iron proteins
Nat. Chem. Biol.
13
147-149
2017
Azotobacter vinelandii
Ivleva, N.B.; Groat, J.; Staub, J.M.; Stephens, M.
Expression of active subunit of nitrogenase via integration into plant organelle genome
PLoS ONE
11
e0160951
2016
Azotobacter vinelandii
Fay, A.W.; Wiig, J.A.; Lee, C.C.; Hu, Y.
Identification and characterization of functional homologs of nitrogenase cofactor biosynthesis protein NifB from methanogens
Proc. Natl. Acad. Sci. USA
112
14829-14833
2015
Azotobacter vinelandii
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