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2013 | 60 | 2 | 217-222
Article title

Single-electron reduction of quinone and nitroaromatic xenobiotics by recombinant rat neuronal nitric oxide synthase

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We examined the kinetics of single-electron reduction of a large number of structurally diverse quinones and nitroaromatic compounds, including a number of antitumour and antiparasitic drugs, and nitroaromatic explosives by recombinant rat neuronal nitric oxide synthase (nNOS, EC, aiming to characterize the role of nNOS in the oxidative stress-type cytotoxicity of the above compounds. The steady-state second-order rate constants (kcat/Km) of reduction of the quinones and nitroaromatics varied from 102 M-1s-1 to 106 M-1s-1, and increased with an increase in their single-electron reduction potentials (E17). The presence of Ca2+/calmodulin enhanced the reactivity of nNOS. These reactions were consistent with an 'outer sphere' electron-transfer mechanism, considering the FMNH./FMNH2 couple of nNOS as the most reactive reduced enzyme form. An analysis of the reactions of nNOS within the 'outer sphere' electron-transfer mechanism gave the approximate values of the distance of electron transfer, 0.39-0.47 nm, which are consistent with the crystal structure of the reductase domain of nNOS. On the other hand, at low oxygen concentrations ([O2] = 40-50 μM), nNOS performs a net two-electron reduction of quinones and nitroaromatics. This implies that NOS may in part be responsible for the bioreductive alkylation by two-electron reduced forms of antitumour aziridinyl-substituted quinones under a modest hypoxia.
Physical description
  • Institute of Biochemistry of Vilnius University, Vilnius, Lithuania
  • Institute of Biochemistry of Vilnius University, Vilnius, Lithuania
  • Institute of Biochemistry of Vilnius University, Vilnius, Lithuania
  • Laboratoire de Chimie & Biochimie Pharmacologiques et Toxicologiques, CNRS UMR 8601, Universite Paris Descartes, 45, Paris, France
  • Laboratoire de Chimie & Biochimie Pharmacologiques et Toxicologiques, CNRS UMR 8601, Universite Paris Descartes, 45, Paris, France
  • Institute of Biochemistry of Vilnius University, Vilnius, Lithuania
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