title: In vitro synthesis of the iron-molybdenum cofactor and maturation of the nif-encoded apodinitrogenase. Effect of substitution of VNFH for NIFH.
abstract: NIFH (the nifH gene product) has several functions in the nitrogenase enzyme system. In addition to reducing dinitrogenase during nitrogenase turnover, NIFH functions in the biosynthesis of the iron-molybdenum cofactor (FeMo-co), and in the processing of alpha2beta2 apodinitrogenase 1 (a catalytically inactive form of dinitrogenase 1 that lacks the FeMo-co) to the FeMo-co-activatable alpha2beta2gamma2 form. The molybdenum-independent nitrogenase 2 (vnf-encoded) has a distinct dinitrogenase reductase protein, VNFH. We investigated the ability of VNFH to function in the in vitro biosynthesis of FeMo-co and in the maturation of apodinitrogenase 1. VNFH can replace NIFH in both the biosynthesis of FeMo-co and in the maturation of apodinitrogenase 1. These results suggest that the dinitrogenase reductase proteins do not specify the heterometal incorporated into the cofactors of the respective nitrogenase enzymes. The specificity for the incorporation of molybdenum into FeMo-co was also examined using the in vitro FeMo-co synthesis assay system.
authors.affiliation: Department of Biochemistry and Center for the Study of Nitrogen Fixation, College of Agricultural and Life Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
authors.forename: R
authors.initials: R
authors.lastname: Chatterjee
authors.forename: R M
authors.initials: RM
authors.lastname: Allen
authors.forename: P W
authors.initials: PW
authors.lastname: Ludden
authors.forename: V K
authors.initials: VK
authors.lastname: Shah
mesh_terms: D001059:Apoproteins / Q000378:metabolism*; D016948:Azotobacter vinelandii / Q000201:enzymology*; D001426:Bacterial Proteins; D002474:Cell-Free System; D008667:Metalloproteins / Q000378:metabolism; D008982:Molybdenum / Q000378:metabolism*; D008983:Molybdoferredoxin / Q000096:biosynthesis*; D009591:Nitrogenase / Q000378:metabolism*; D010088:Oxidoreductases
chemical_list: D001059:Apoproteins; D001426:Bacterial Proteins; D008667:Metalloproteins; D008983:Molybdoferredoxin; D008982:Molybdenum; D010088:Oxidoreductases; C097905:vnfDGK protein, Azotobacter; D009591:Nitrogenase; C029130:nitrogenase reductase