Cryoradiolysis and cryospectroscopy for studies of heme-oxygen intermediates in cytochromes P450

I. G. Denisov, Y. V. Grinkova, S. G. Sligar

Research output: Chapter in Book/Report/Conference proceedingChapter


Cryogenic radiolytic reduction is one of the most straightforward and convenient methods of generation and stabilization of reactive iron-oxygen intermediates for mechanistic studies in chemistry and biochemistry. The method is based on one-electron reduction of the precursor complex in frozen solution via exposure to the ionizing radiation at cryogenic temperatures. Such approach allows for accumulation of the fleeting reactive complexes which otherwise could not be generated at sufficient amount for structural and mechanistic studies. Application of this method allowed for characterizing of peroxo-ferric and hydroperoxo-ferric intermediates, which are common for the oxygen activation mechanism in cytochromes P450, heme oxygenases, and nitric oxide synthases, as well as for the peroxide metabolism by peroxidases and catalases.

Original languageEnglish (US)
Title of host publicationSpectroscopic Methods of Analysis
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Number of pages17
ISBN (Print)9781617798054
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Cytochrome P450
  • Hydroperoxo-ferric complex
  • Low-temperature methods
  • Peroxo-ferric complex
  • Radiolysis
  • Radiolytic reduction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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