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 | 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 | Comment | Organism | Structure |
---|---|---|---|
CO | binds to the active site | Azotobacter vinelandii |
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 | 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 | 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 | UniProt | Comment | Textmining |
---|---|---|---|
Azotobacter vinelandii | - |
- |
- |
Azotobacter vinelandii DJ1310 | - |
- |
- |
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 | 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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
2 | - |
wild-type enzyme, C2H2 reduction | Azotobacter vinelandii |
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 Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Azotobacter vinelandii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Azotobacter vinelandii |
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 |