Fe protein cycle
The nitrogenase Fe protein serves to deliver electrons, one at a time, to the MoFe protein in a process coupled to the hydrolysis of two MgATP molecules (28, 31). The Fe protein is a homodimer (Mr≈ 64,000) having one nucleotide (MgATP/MgADP) binding site buried within each subunit and a single [4Fe-4S] cluster bridging the two subunits (Figure 1) (13). The participation of the Fe protein in the nitrogenase mechanism can be considered to operate in a three state cycle (called the Fe protein cycle) (Figure 2, top). In this cycle, the reduced Fe protein, with its [4Fe-4S] cluster in the 1+ oxidation state, has two bound MgATP molecules. It is this state that transiently associates with the MoFe protein (32). During this association, the two MgATP molecules are hydrolyzed to two MgADP molecules, and a single electron is transferred from the Fe protein [4Fe-4S] cluster into the MoFe protein. The oxidized Fe protein ([4Fe-4S]2+), now with two bound MgADP molecules, then dissociates from the MoFe protein in what is the overall rate-limiting step for nitrogenase catalysis (33). The released Fe protein is then regenerated in two steps. The MgADP molecules are replaced by MgATP, and the [4Fe-4S]2+ cluster is reduced to the 1+ oxidation state, with the order of these two events being unclear (34). The physiological reductant for the Fe protein depends on the organism, with reduced ferredoxin or flavodoxin being the most common immediate electron donor (35, 36).