Literature summary extracted from
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 (2010), J. Inorg. Biochem., 104, 385-389.
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
1.18.6.1 |
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 |
Azotobacter vinelandii |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.18.6.1 |
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 |
Azotobacter vinelandii |
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
1.18.6.1 |
Iron |
in FeMo cofactor and MoFe protein |
Azotobacter vinelandii |
|
1.18.6.1 |
Molybdenum |
in FeMo cofactor and MoFe protein |
Azotobacter vinelandii |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.18.6.1 |
Azotobacter vinelandii |
P07328 |
- |
- |
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 |
2.2 |
- |
about, micromol/min/mg MoFe protein, mutant enzyme, pH not specified in the publication, temperature not specified in the publication |
Azotobacter vinelandii |
1.18.6.1 |
2.316 |
- |
about, micromol/min/mg MoFe protein, wild-type enzyme, pH not specified in the publication, temperature not specified in the publication |
Azotobacter vinelandii |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.18.6.1 |
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 |
Azotobacter vinelandii |
? |
- |
? |
|
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.18.6.1 |
iron-molybdenum cofactor |
modelling of the FeMo-cofactor binding site in the alpha70Ile MoFe protein structure |
Azotobacter vinelandii |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
1.18.6.1 |
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 |
Azotobacter vinelandii |