Complex formation and O2 sensitivity of Azotobacter vinelandii nitrogenase and its component proteins.
The O2 stability of the MoFe protein, the Fe protein, a 1:1 mixture of these proteins, and a 1:1 mixture in the presence of the Azotobacter vinelandii FeS-II protein has been studied as a function of time under controlled O2 partial pressures. The Fe protein is much more sensitive to O2 exposure than is the MoFe protein. The presence of the FeS-II protein at a 1:1 ratio with the component proteins measurably increases the O2 stability of the MoFe and Fe proteins. O2 inactivation of the MoFe protein was studied in some detail and found to be quite complex. At least three partially overlapping reactions are suggested. The first is the reversible oxidation of the metal clusters of the MoFe protein to the combined extent of 12 electrons with full retention of activity. The second phase consists primarily of activity loss with little increase in the extent of reversible oxidation. The third phase continues to decrease the protein activity but is also accompanied by formation of a g = 2.0 EPR signal and more extensive oxidation. Ultracentrifugation studies of the FeS-II protein at a 1:1:1 ratio with the Fe and MoFe proteins do not support the formation of the Bulen complex. The formation of other O2-stable complexes is discussed.
Wang
Z C
ZC
Burns
A
A
Watt
G D
GD
D001395:Azotobacter / Q000201:enzymology*; D004355:Drug Stability; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D046911:Macromolecular Substances; D009591:Nitrogenase / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D011485:Protein Binding
D046911:Macromolecular Substances; D009591:Nitrogenase; D010100:Oxygen