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Literature summary extracted from

  • 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 (2003), Biochemistry, 42, 9102-9109.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.18.6.1 expression of His-tagged wild-type and V70A mutant MoFe protein in strain DJ1310, expression of the wild-type Fe protein Azotobacter vinelandii

Protein Variants

EC Number Protein Variants Comment Organism
1.18.6.1 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 Azotobacter vinelandii

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.18.6.1 CO binds to the active site Azotobacter vinelandii

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.18.6.1 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 Azotobacter vinelandii

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.18.6.1 Iron dependent on, the enzyme complex contains a molybdenum-iron protein harboring the active site, the enzyme complex contains also a dimeric iron protein Azotobacter vinelandii
1.18.6.1 Mg2+
-
 Azotobacter vinelandii
1.18.6.1 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 Azotobacter vinelandii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.18.6.1 reduced ferredoxin + H+ + N2 + ATP + H2O Azotobacter vinelandii
-
 oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
 ir
1.18.6.1 reduced ferredoxin + H+ + N2 + ATP + H2O Azotobacter vinelandii DJ1310
-
 oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
 ir

Organism

EC Number Organism UniProt Comment Textmining
1.18.6.1 Azotobacter vinelandii
-
-
-
1.18.6.1 Azotobacter vinelandii DJ1310
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.18.6.1 recombinant wild-type Fe protein, recombinant His-tagged wild-type MoFe protein and recombinant His-tagged MoFe protein mutant V70A, to homogeneity Azotobacter vinelandii

Reaction

EC Number Reaction Comment Organism Reaction ID
1.18.6.1 4 reduced ferredoxin + 8 H+ + N2 + 16 ATP + 16 H2O = 4 oxidized ferredoxin + H2 + 2 NH3 + 16 ADP + 16 phosphate active site is located on the MoFe cofactor involving residues alphaR96, alphaG69, alphaV70, and alphaH195 Azotobacter vinelandii
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.18.6.1 2
-
 wild-type enzyme, C2H2 reduction Azotobacter vinelandii

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.18.6.1 C2H2 + ?
-
 Azotobacter vinelandii ?
-
 ?
1.18.6.1 C2H2 + ?
-
 Azotobacter vinelandii DJ1310 ?
-
 ?
1.18.6.1 propargyl alcohol + ? wild-type enzyme and V70A mutant MoFe protein-containing enzyme Azotobacter vinelandii ?
-
 ?
1.18.6.1 propargyl alcohol + ? wild-type enzyme and V70A mutant MoFe protein-containing enzyme Azotobacter vinelandii DJ1310 ?
-
 ?
1.18.6.1 reduced ferredoxin + H+ + N2 + ATP + H2O
-
 Azotobacter vinelandii oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
 ir
1.18.6.1 reduced ferredoxin + H+ + N2 + ATP + H2O
-
 Azotobacter vinelandii DJ1310 oxidized ferredoxin + H2 + NH3 + ADP + phosphate
-
 ir

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.18.6.1 25
-
 assay at Azotobacter vinelandii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.18.6.1 7
-
 assay at Azotobacter vinelandii

Cofactor

EC Number Cofactor Comment Organism Structure
1.18.6.1 Ferredoxin
-
 Azotobacter vinelandii
1.18.6.1 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 Azotobacter vinelandii