title: Ferredoxin-NADP(+) reductase uses the same site for the interaction with ferredoxin and flavodoxin.
abstract: The enzyme ferredoxin-NADP(+) reductase (FNR) forms a 1 : 1 complex with ferredoxin (Fd) or flavodoxin (Fld) that is stabilised by both electrostatic and hydrophobic interactions. The electrostatic interactions occur between acidic residues of the electron transfer (ET) protein and basic residues on the FNR surface. In the present study, several charge-reversal mutants of FNR have been prepared at the proposed site of interaction of the ET protein: R16E, K72E, K75E, K138E, R264E, K290E and K294E. All of these mutants have been assayed for reactivity with Fd and Fld using steady-state and stopped-flow kinetics. Their abilities for complex formation with the ET proteins have also been tested. The data presented here indicate that the mutated residues situated within the FNR FAD-binding domain are more important for achieving maximal ET rates, either with Fd or Fld, than those situated within the NADP(+)-binding domain, and that both ET proteins occupy the same region for the interaction with the reductase. In addition, each individual residue does not appear to participate to the same extent in the different processes with Fd and Fld.
authors.affiliation: Departamento de Bioquímica y Biología Molecular y Celular Facultad de Ciencias, Universidad de Zaragoza E-50009 Saragossa, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Martínez-Júlvez
authors.forename: M
authors.initials: M
authors.lastname: Medina
authors.forename: C
authors.initials: C
authors.lastname: Gómez-Moreno
mesh_terms: D017033:Anabaena / Q000201:enzymology; D004926:Escherichia coli; D005287:Ferredoxin-NADP Reductase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005288:Ferredoxins / Q000378:metabolism*; D005418:Flavodoxin / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed
chemical_list: D005288:Ferredoxins; D005418:Flavodoxin; D005287:Ferredoxin-NADP Reductase