NifI inhibits nitrogenase by competing with Fe protein for binding to the MoFe protein.
Reduction of substrate by nitrogenase requires direct electron transfer from the Fe protein to the MoFe protein. Inhibition of nitrogenase activity in Methanococcus maripaludis occurs when the regulatory protein NifI(1,2) binds the MoFe protein. This inhibition is relieved by 2-oxoglutarate. Here we present evidence that NifI(1,2) binding prevents association of the two nitrogenase components. Increasing amounts of Fe protein competed with NifI(1,2), decreasing its inhibitory effect. NifI(1,2) prevented the co-purification of MoFe protein with a mutant form of the Fe protein that forms a stable complex with the MoFe protein, and NifI(1,2) was unable to bind to an AlF4--stabilized Fe protein:MoFe protein complex. NifI(1,2) inhibited ATP- and MoFe protein-dependent oxidation of the Fe protein, and 2OG relieved this inhibition. These results support a model where NifI(1,2) competes with the Fe protein for binding to MoFe protein and prevents electron transfer.
Dodsworth
Jeremy A
JA
Department of Microbiology, University of Washington, P.O. Box 357242, Seattle, WA 98195-7242, USA.
Leigh
John A
JA
D000255:Adenosine Triphosphate / Q000737:chemistry; D019843:Archaeal Proteins / Q000737:chemistry*; D001667:Binding, Competitive; D017017:Methanococcus / Q000378:metabolism*; D008983:Molybdoferredoxin / Q000737:chemistry*; D009586:Nitrogen Fixation; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry*; D051922:PII Nitrogen Regulatory Proteins / Q000737:chemistry*; D011485:Protein Binding
D019843:Archaeal Proteins; D008983:Molybdoferredoxin; C423625:NifI1 protein, Methanococcus maripaludis; C423626:NifI2 protein, Methanococcus maripaludis; D051922:PII Nitrogen Regulatory Proteins; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; C029130:nitrogenase reductase