Roles of phenylalanine at position 120 and glutamic acid at position 222 in the oxidation of chiral substrates by cytochrome P450 2D6

Kazufumi Masuda, Keietsu Tamagake, Takashi Katsu, Fumihiro Torigoe, Keita Saito, Nobumitsu Hanioka, Shigeru Yamano, Shigeo Yamamoto, Shizuo Narimatsu

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The roles of Phe-120 and Glu-222 in the oxidation of chiral substrates bunitrolol (BTL) and bufuralol (BF) by CYP2D6 are discussed. Wild-type CYP2D6 (CYP2D6-WT) oxidized BTL to 4-hydroxybunitrolol (4-OH-BTL) with substrate enantioselectivity of (R)-(+)-BTL > (S)-(-)-BTL. The same enzyme converted BF into 1″-hydroxybufuralol with substrate enantioselectivity of (R)-BF ≫ (S)-BF and metabolite diastereoselectivity of (1″ R)-OH < (1″ S)-OH. The substitution of Phe-120 by alanine markedly increased the apparent Km and Vmax values for enantiomeric BTL 4-hydroxylation by CYP2D6. In contrast, the same substitution caused an increase only in V max values of (5)-BF 1″-hydroxylation without changing apparent Km values, while kinetic parameters (Km, and V max values) for (R)-BF 1″-hydroxylation remained unchanged. Furthermore, the substitution of Glu-222 as well as Glu-216 by alanine remarkably decreased both the apparent Km and Vmax values without changing substrate enantioselectivity or metabolite diastereoselectivity. A computer-assisted simulation study using energy minimization and molecular dynamics techniques indicated that the hydrophobic interaction of an aromatic moiety of the substrate with Phe-120 and the ionic interaction of a basic nitrogen atom of the substrate with Glu-222 in combination with Glu-216 play important roles in the binding of BF and BTL by CYP2D6 and the orientation of these substrates in the active-site cavity. This modeling yielded a convincing explanation for the reversal of substrate enantioselectivity in BTL 4-hydroxylation between CYP2D6-WT and CYP2D6-V374M having methionine in place of Val-374, which supports the validity of this modeling.

Original languageEnglish
Pages (from-to)167-176
Number of pages10
JournalChirality
Volume18
Issue number3
DOIs
Publication statusPublished - 2006
Externally publishedYes

Keywords

  • Bufuralol
  • Bunitrolol
  • CYP2D6
  • Glu-216
  • Glu-222
  • Met-374
  • Phe-120
  • Substrate enantioselectivity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Catalysis
  • Pharmacology
  • Drug Discovery
  • Spectroscopy
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Roles of phenylalanine at position 120 and glutamic acid at position 222 in the oxidation of chiral substrates by cytochrome P450 2D6'. Together they form a unique fingerprint.

  • Cite this

    Masuda, K., Tamagake, K., Katsu, T., Torigoe, F., Saito, K., Hanioka, N., Yamano, S., Yamamoto, S., & Narimatsu, S. (2006). Roles of phenylalanine at position 120 and glutamic acid at position 222 in the oxidation of chiral substrates by cytochrome P450 2D6. Chirality, 18(3), 167-176. https://doi.org/10.1002/chir.20246