Cryoradiolysis and cryospectroscopy for studies of heme-oxygen intermediates in cytochromes p450.

Methods in Molecular Biology
I G DenisovS G Sligar

Abstract

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.

Citations

Dec 17, 2015·Proceedings of the National Academy of Sciences of the United States of America·Piotr J MakJames R Kincaid
Feb 9, 2017·Chemical Reviews·Ilia G Denisov, Stephen G Sligar
Jul 3, 2017·Biochimica Et Biophysica Acta. Proteins and Proteomics·Piotr J Mak, Ilia G Denisov

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