EPR and redox properties of periplasmic nitrate reductase from Desulfovibrio desulfuricans ATCC 27774

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Abstract

Nitrate reductases are enzymes that catalyze the conversion of nitrate to nitrite. We report here electron paramagnetic resonance (EPR) studies in the periplasmic nitrate reductase isolated from the sulfate-reducing bacteria Desulfovibrio desulfuricans ATCC 27774. This protein, belonging to the dimethyl sulfoxide reductase family of mononuclear Mo-containing enzymes, comprises a single 80-kDa subunit and contains a Mo bis(molybdopterin guanosine dinucleotide) cofactor and a [4Fe-4S] cluster. EPR-monitored redox titrations, carried out with and without nitrate in the potential range from 200 to -500 mV, and EPR studies of the enzyme, in both catalytic and inhibited conditions, reveal distinct types of Mo(V) EPR-active species, which indicates that the Mo site presents high coordination flexibility. These studies show that nitrate modulates the redox properties of the Mo active site, but not those of the [4Fe-4S] center. The possible structures and the role in catalysis of the distinct Mo(V) species detected by EPR are discussed.

Original languageEnglish
Pages (from-to)609-616
Number of pages8
JournalJBIC Journal of Biological Inorganic Chemistry
Volume11
Issue number5
DOIs
Publication statusPublished - 1 Jul 2006

Keywords

  • Dimethyl sulfoxide reductase family
  • Electron paramagnetic resonance
  • Molybdenum-containing enzymes
  • Periplasmic nitrate reductase
  • Redox titration

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