Electron transport to nitrogenase in Klebsiella pneumoniae: purification and properties of the nifJ protein

Abstract

In Klebsiella pneumoniae, the physiological electron flow to nitrogenase involves specifically, in addition to nitrogenase reductase, the products of the nifF and nifJ genes. The J protein was purified to homogeneity and was found to be an iron-sulfur protein devoid of molybdenum. In its native state, the J protein is a dimer of Mr about 245 000, made up of two subunits of the same molecular weight. It contains about 30 mol iron and 24 mol labile sulfur/mol protein. The addition of J protein to crude extracts of a nifJ mutant reestablishes pyruvate-supported acetylene-reducing activity. This activity is further enhanced by addition of pure nitrogenase (Kp1). Based on its physical properties, the J protein is probably an oxidoreductase whose physiological role might be to transfer electrons from a metabolic donor to the F protein. In addition, another protein whose activity is also dependent on the nifJ gene seems to be required for the formation of a fully active Kp1.