ApoNifH functions in iron-molybdenum cofactor synthesis and apodinitrogenase maturation.
NifH (dinitrogenase reductase) has three important roles in the nitrogenase enzyme system. In addition to its role as the obligate electron donor to dinitrogenase, NifH is required for the iron-molybdenum cofactor (FeMo-co) synthesis and apodinitrogenase maturation. We have investigated the requirement of the Fe-S cluster of NifH for these processes by preparing apoNifH. The 4Fe-4S cluster of NifH was removed by chelation of the cluster with alpha, alpha'-bipyridyl. The resulting apoNifH was tested in in vitro FeMo-co synthesis and apodinitrogenase maturation reactions and was found to function in both these processes. Thus, the presence of a redox active 4Fe-4S cluster in NifH is not required for its function in FeMo-co synthesis and in apodinitrogenase maturation. This, in turn, implies that the role of NifH in these processes is not one of electron transfer or of iron or sulfur donation.
Rangaraj
P
P
Department of Biochemistry and Center for the Study of Nitrogen Fixation, College of Agricultural and Life Sciences, University of Wisconsin, Madison, WI 53706, USA.
Shah
V K
VK
Ludden
P W
PW
D000255:Adenosine Triphosphate / Q000378:metabolism; D000693:Anaerobiosis; D001051:Apoenzymes / Q000378:metabolism*; D016948:Azotobacter vinelandii / Q000201:enzymology*; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000302:isolation & purification / Q000378:metabolism*; D007506:Iron-Sulfur Proteins / Q000378:metabolism; D007700:Kinetics; D046911:Macromolecular Substances; D008983:Molybdoferredoxin / Q000096:biosynthesis*; D009591:Nitrogenase / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D010088:Oxidoreductases
D001051:Apoenzymes; D004792:Enzyme Precursors; D007506:Iron-Sulfur Proteins; D046911:Macromolecular Substances; D008983:Molybdoferredoxin; D000255:Adenosine Triphosphate; D010088:Oxidoreductases; D009591:Nitrogenase; C029130:nitrogenase reductase