Electron-paramagnetic-resonance studies on the redox properties of the molybdenum-iron protein of nitrogenase between +50 and -450 mV.

The midpoint potentials, Em, for the oxidation of the characteristic e.p.r. signal with g values near 4.3, 3.7 and 2.01, of the nitrogenase Mo-Fe proteins from a number of bacteria were measured. They were 0mV for Clostridium pasteurianum, -42mV for Azotobacter chroococcum and Azotobacter vinelandii, -95mV for Bacillus polymyxa and -180mV for Klebsiella pneumoniae Mo-Fe proteins at pH 7.9. The oxidations were thermodynamically reversible for the proteins from A. chroococcum, A. vinelandii and K. pneumoniae and the Em was independent of protein activity for this last protein. The protein from C. pasteurianum required a lower potential for reduction than for oxidation, and the oxidation of the protein from B. polymyxa was only 70% reversible. The apparent Em of the latter protein was decreased by 40mV in the presence of 60mM-MgCl2. The pH-dependence of the Em of the protein from K. pneumoniae was interpreted in terms of a single ionization, not directly associated with the e.p.r.-active centre, with a pKa of 7.0 in the oxidized form of the protein and a pH-independent region at low pH (Em = 118 +/- 6.3 mV). Approx. 20% increase in activity after oxidation was observed for the proteins from B. polymyxa, A. chroococcum and K. pneumoniae. The significance of the above results and their relationship to other published data are discussed.