Interaction of nitric oxide with photoexcited rose bengal: Evidence for one-electron reduction of nitric oxide to nitroxyl anion

Ravinder Jit Singh, Neil Hogg, B. Kalyanaraman

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8 Scopus citations


The interaction of nitric oxide (·NO) with Rose Bengal (RB) in the presence of electron donors was investigated. Upon illumination of a mixture of RB and·NO with visible light, an enhancement in the rate of·NO consumption was observed that increased with increasing RB concentration. In the presence of electron donors (NADH, glutathione, or ascorbate), the rates of·NO depletion increased further. NADH enhanced·NO depletion to a greater extent than either glutathione or ascorbate. Photoactivated RB under anaerobic conditions reacts with NADH to form the RB anion radical (RB·—), which has a characteristic visible absorption band centered at 418 nm. Rose Bengal anion radical disproportionates to give RB and a colorless reduced form of RB, RBH—. The net result of this process is the photobleaching of RB. The presence of·NO during irradiation of RB and NADH introduced a lag time into the kinetics of RB photobleaching. The length of this lag time was proportional to the concentration of·NO. A similar lag time, which was also dependent on the·NO concentration, was observed in the kinetics of formation of RB·—. The three-line electron spin resonance (ESR) spectrum of RB·—, with an intensity ratio 1:2:1, was obtained during irradiation of RB and NADH under anaerobic conditions.·NO introduced a concentration-dependent lag time into the kinetics of the appearance of this ESR signal. We propose that·NO oxidizes RB·—to regenerate RB and thus inhibit photobleaching until·NO is consumed. This reaction predicts the formation of NO—, the one-electron reduced form of·NO. Nitrous oxide, a characteristic dimerization product of NO—, was detected by gas chromatography. This evidence indicates the occurrence of a Type I mechanism between photoactivated RB and·NO.

Original languageEnglish (US)
Pages (from-to)367-373
Number of pages7
JournalArchives of Biochemistry and Biophysics
Issue number2
StatePublished - Dec 20 1995



  • Nitric oxide
  • Nitrous oxide
  • Nitroxyl anion
  • Photosensitization
  • Rose Bengal

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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