Revisiting the mechanism of P450 enzymes with the radical clocks norcarane and spiro[2,5]octane

Karine Auclair, Zhengbo Hu, Dorothy M. Little, Paul R. Ortiz de Montellano, John Taylor Groves

Research output: Contribution to journalArticle

132 Scopus citations

Abstract

Norcarane (1) and spiro[2.5]octane (2) yield different product distributions depending on whether they are oxidized via concerted, radical, or cationic mechanisms. For this reason, these two probes were used to investigate the mechanisms of hydrocarbon hydroxylation by two mammalian and two bacterial cytochrome P450 enzymes. Products indicative of a radical intermediate with a lifetime ranging from 16 to 52 ps were detected during the oxidation of norcarane by P450cam (CYP101), P450BM3 (CYP102), CYP2B1, and CYP2E1. Trace amounts of the cation rearrangement product were observed with norcarane for all but CYP2E1, while no cation or radical rearrangement products were observed for spiro[2.5]octane. The results for the oxidation of norcarane with a radical rearrangement rate of 2 × 108 s-1 are consistent with the involvement of a two-state radical rebound mechanism, while for the slower (5 × 107 s-1) spiro[2,5]-oct-4-yl radical rearrangement products were beyond detection. Taken together with earlier data for the hydroxylation of bicyclo[2.1.0]pentane, which also suggested a 50 ps radical lifetime, these three structurally similar and functionally simple substrates show a consistent pattern of rearrangement that supports a radical rebound mechanism for this set of cytochrome P450 enzymes.

Original languageEnglish (US)
Pages (from-to)6020-6027
Number of pages8
JournalJournal of the American Chemical Society
Volume124
Issue number21
DOIs
StatePublished - May 20 2002

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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