The oxidant-responsive diaphorase of Rhodobacter capsulatus is a ferredoxin (flavodoxin)-NADP(H) reductase.
Challenge of Rhodobacter capsulatus cells with the superoxide propagator methyl viologen resulted in the induction of a diaphorase activity identified as a member of the ferredoxin (flavodoxin)-(reduced) nicotinamide adenine dinucleotide phosphate (NADP(H)) reductase (FPR) family by N-terminal sequencing. The gene coding for Rhodobacter FPR was cloned and expressed in Escherichia coli. Both native and recombinant forms of the enzyme were purified to homogeneity rendering monomeric products of approximately 30 kDa with essentially the same spectroscopic and kinetic properties. They were able to bind and reduce Rhodobacter flavodoxin (NifF) and to mediate typical FPR activities such as the NADPH-driven diaphorase and cytochrome c reductase.
Instituto de Biología Molecular y Celular de Rosario, Universidad Nacional de Rosario and CONICET, Suipacha 531, S2002LRK Rosario, Argentina.
Cristian
C
Bittel
Leandro C
LC
Tabares
Martín
M
Armesto
Néstor
N
Carrillo
Néstor
N
Cortez
D003001:Cloning, Molecular; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D005287:Ferredoxin-NADP Reductase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005418:Flavodoxin / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D016680:Genome, Bacterial; D009245:NADH Dehydrogenase / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016354:Rhodobacter capsulatus / Q000201:enzymology* / Q000235:genetics; D013053:Spectrophotometry / Q000379:methods
D005418:Flavodoxin; D011994:Recombinant Proteins; D009249:NADP; D005287:Ferredoxin-NADP Reductase; D009245:NADH Dehydrogenase; D008058:Dihydrolipoamide Dehydrogenase