Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.18.6.1 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)

Cloned (Comment) Organism
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

Protein Variants Comment Organism
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

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
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

Metals/Ions Comment Organism Structure
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
Mg2+
-
Azotobacter vinelandii
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)

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

Organism

Organism UniProt Comment Textmining
Azotobacter vinelandii
-
-
-
Azotobacter vinelandii DJ1310
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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

Reaction Comment Organism Reaction ID
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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
Ferredoxin
-
Azotobacter vinelandii
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