Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cloning, Molecular
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Dihydrolipoamide Dehydrogenase / genetics
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Dihydrolipoamide Dehydrogenase / isolation & purification
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Dihydrolipoamide Dehydrogenase / metabolism*
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Escherichia coli / metabolism
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Ferredoxin-NADP Reductase / genetics
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Ferredoxin-NADP Reductase / isolation & purification
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Ferredoxin-NADP Reductase / metabolism*
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Flavodoxin / metabolism
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Gene Expression Regulation, Bacterial
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Genome, Bacterial
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NADH Dehydrogenase / metabolism
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NADP / metabolism
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Oxidative Stress / physiology
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Rhodobacter capsulatus / enzymology*
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Rhodobacter capsulatus / genetics
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Spectrophotometry / methods
Substances
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Flavodoxin
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Recombinant Proteins
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NADP
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Ferredoxin-NADP Reductase
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NADH Dehydrogenase
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Dihydrolipoamide Dehydrogenase