Catalytic mechanism of cytochrome P450 for 5′-hydroxylation of nicotine: Fundamental reaction pathways and stereoselectivity

Dongmei Li, Xiaoqin Huang, Keli Han, Chang Guo Zhan

Research output: Contribution to journalArticlepeer-review

114 Scopus citations

Abstract

A series of computational methods were used to study how cytochrome P450 2A6 (CYP2A6) interacts with (S)-(-)-nicotine, demonstrating that the dominant molecular species of (S)-(-)-nicotine in CYP2A6 active site exists in the free base state (with two conformations, SRt and SRc), despite the fact that the protonated state is dominant for the free ligand in solution. The computational results reveal that the dominant pathway of nicotine metabolism in CYP2A6 is through nicotine free base oxidation. Further, first-principles quantum mechanical/molecular mechanical free energy (QM/MM-FE) calculations were carried out to uncover the detailed reaction pathways for the CYP2A6-catalyzed nicotine 5′-hydroxylation reaction. In the determined CYP2A6-(S)-(-)-nicotine binding structures, the oxygen of Compound I (Cpd I) can abstract a hydrogen from either the trans-5′-or the cis-5′-position of (S)-(-)-nicotine. CYP2A6-catalyzed (S)-(-)-nicotine 5′-hydroxylation consists of two reaction steps, that is, the hydrogen transfer from the 5′-position of (S)-(-)-nicotine to the oxygen of Cpd I (the H-transfer step), followed by the recombination of the (S)-(-)-nicotine moiety with the iron-bound hydroxyl group to generate the 5′-hydroxynicotine product (the O-rebound step). The H-transfer step is rate-determining. The 5′-hydroxylation proceeds mainly with the stereoselective loss of the trans-5′-hydrogen, that is, the 5′-hydrogen trans to the pyridine ring. The calculated overall stereoselectivity of ∼97% favoring the trans-5′-hydroxylation is close to the observed stereoselectivity of 89-94%. This is the first time it has been demonstrated that a CYP substrate exists dominantly in one protonation state (cationic species) in solution, but uses its less-favorable protonation state (neutral free base) to perform the enzymatic reaction.

Original languageEnglish
Pages (from-to)7416-7427
Number of pages12
JournalJournal of the American Chemical Society
Volume133
Issue number19
DOIs
StatePublished - May 18 2011

ASJC Scopus subject areas

  • Catalysis
  • General Chemistry
  • Biochemistry
  • Colloid and Surface Chemistry

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