J Integr Plant Biol. ›› 1994, Vol. 36 ›› Issue (6): -.

• Research Articles •    

The Effect of Manganese on the Reconstitution of Partial Metallocluster-deficient Nitrogenase Molybdenum-Iron Protein

Huang Ju-fu, Wang Zhi-ping, Luo Ai-ling, Zhong Ze-pu and Li Jia-ge   

Abstract: By treating the reduced MoFe protein of nitrogenase from Azotobacter vinelandii with O-phenanthroline (O-phen) and O2, inactive MoFe protein which was partialy deficient in both P-cluster and FeMoco could be obtained. After incubating the inactive protein with a reconstituent solution containing KMnO4, ferric homocitrate, Na2S and dithiothreitol, a reconstituted protein could be obtained. The absorption spectrum and C2H2, H+ and N2 reduction activity of the reconstituted protein could be well restored to the state of the reduced MoFe protein. However, the α-helix and CD spectrum at 380—550 nm and at 620—670 nm of the reconstituted protein were somewhat different from those of the reduced MoFe protein. The results showed that: (1) the reconstituted protein was composed of the assembled protein which might be a MnFe protein due to the reconstitution of the metalloclusterdeficient MoFe protein with Mn-containing solution and MoFe protein in which metalloclusters were still intact after the treatment with O-phen and O2; (2) It might be possible that the MnFe protein and MoFe protein were similar in the ability of nitrogen fixation, but were somewhat different in the structure from each other.

Key words: Assembly in vitro, Manganese-iron protein, Molybdenum-iron protein, Nitrogenase, Azotobacter vinelandii

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