Enantioselectivity of bunitrolol 4-hydroxylation is reversed by the change of an amino acid residue from valine to methionine at position 374 of cytochrome P450-2D6

Shizuo Narimatsu, Rika Kato, Toshiharu Horie, Satoshi Ono, Michio Tsutsui, Yoshiyasu Yabusaki, Shigeru Ohmori, Mitsukazu Kitada, Takao Ichioka, Noriaki Shimada, Ryuichi Kato, Tsutomu Ishikawa

Research output: Contribution to journalArticle

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Abstract

The enantioselectivity of 4-hydroxylation of bunitrolol (BTL), a β- adrenoceptor blocking drug, was studied in microsomes from human liver, human hepatoma (Hep G2) cells expressing CYP2D6, and lymphoblastoid cells expressing CYP2D6. Kinetics in human liver microsomes showed that the Vmax value for (+)-BTL was 2.1-fold that of (-)-BTL, and that the Km value for (+)-BTL was lower than that for the (-)antipode, resulting in the intrinsic clearance (Vmax/Km) of (+)-BTL being 2.1-fold over its (-)-antipode. CYP2D6 (CYP2D6-met) expressed in Hep G2 cells had a methionine residue at position 373 of the amino acid sequence and a rat-type N-terminal peptide (MELLNGTGLWSM) instead of the human-type (MGLEALVPLAVIV), and showed enantioselectivity of [(+)-BTL <(-)-BTL] for the rate of BTL 4- hydroxylation. In contrast, enantioselectivity [(+)-BTL > (-)-BTL] for Hep G2-CYP2D6 (CYP2D6-val) with a human-type N-terminal peptide that had a valine residue at 374, which corresponds to the methionine of the CYP2D6-met variant, was the same as that for human liver microsomes. We further confirmed that CYP2D6-met and CYP2D6-val expressed in human lymphoblastoid cells, both of which have methionine and valine, respectively, at position 374 and a human-type N-terminal peptide, exhibited the same enantioselectivities as those obtained from CYP2D6-met and CYP2D6-val expressed in the Hep G2 cell system. These results indicate that the amino acid at 374 of CYP2D6 is one of the key factors influencing the enantioselectivity of BTL 4-hydroxylation.

Original languageEnglish
Pages (from-to)1-9
Number of pages9
JournalChirality
Volume11
Issue number1
DOIs
Publication statusPublished - 1999

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Cytochrome P-450 CYP2D6
Hydroxylation
Enantioselectivity
Valine
Methionine
Amino acids
Liver
Peptides
Amino Acids
Cells
Hep G2 Cells
Liver Microsomes
Rats
Kinetics
bunitrolol
Cytochrome P-450 Enzyme System
Adrenergic Receptors
Amino Acid Sequence
Hepatocellular Carcinoma

Keywords

  • Amino acid
  • Bunitrolol 4-hydroxylation
  • CYP2D6- valine
  • CYP2D6-methionine
  • Enantioselectivity

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Organic Chemistry
  • Pharmacology

Cite this

Enantioselectivity of bunitrolol 4-hydroxylation is reversed by the change of an amino acid residue from valine to methionine at position 374 of cytochrome P450-2D6. / Narimatsu, Shizuo; Kato, Rika; Horie, Toshiharu; Ono, Satoshi; Tsutsui, Michio; Yabusaki, Yoshiyasu; Ohmori, Shigeru; Kitada, Mitsukazu; Ichioka, Takao; Shimada, Noriaki; Kato, Ryuichi; Ishikawa, Tsutomu.

In: Chirality, Vol. 11, No. 1, 1999, p. 1-9.

Research output: Contribution to journalArticle

Narimatsu, S, Kato, R, Horie, T, Ono, S, Tsutsui, M, Yabusaki, Y, Ohmori, S, Kitada, M, Ichioka, T, Shimada, N, Kato, R & Ishikawa, T 1999, 'Enantioselectivity of bunitrolol 4-hydroxylation is reversed by the change of an amino acid residue from valine to methionine at position 374 of cytochrome P450-2D6', Chirality, vol. 11, no. 1, pp. 1-9. https://doi.org/10.1002/(SICI)1520-636X(1999)11:1<1::AID-CHIR1>3.0.CO;2-E
Narimatsu, Shizuo ; Kato, Rika ; Horie, Toshiharu ; Ono, Satoshi ; Tsutsui, Michio ; Yabusaki, Yoshiyasu ; Ohmori, Shigeru ; Kitada, Mitsukazu ; Ichioka, Takao ; Shimada, Noriaki ; Kato, Ryuichi ; Ishikawa, Tsutomu. / Enantioselectivity of bunitrolol 4-hydroxylation is reversed by the change of an amino acid residue from valine to methionine at position 374 of cytochrome P450-2D6. In: Chirality. 1999 ; Vol. 11, No. 1. pp. 1-9.
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abstract = "The enantioselectivity of 4-hydroxylation of bunitrolol (BTL), a β- adrenoceptor blocking drug, was studied in microsomes from human liver, human hepatoma (Hep G2) cells expressing CYP2D6, and lymphoblastoid cells expressing CYP2D6. Kinetics in human liver microsomes showed that the Vmax value for (+)-BTL was 2.1-fold that of (-)-BTL, and that the Km value for (+)-BTL was lower than that for the (-)antipode, resulting in the intrinsic clearance (Vmax/Km) of (+)-BTL being 2.1-fold over its (-)-antipode. CYP2D6 (CYP2D6-met) expressed in Hep G2 cells had a methionine residue at position 373 of the amino acid sequence and a rat-type N-terminal peptide (MELLNGTGLWSM) instead of the human-type (MGLEALVPLAVIV), and showed enantioselectivity of [(+)-BTL <(-)-BTL] for the rate of BTL 4- hydroxylation. In contrast, enantioselectivity [(+)-BTL > (-)-BTL] for Hep G2-CYP2D6 (CYP2D6-val) with a human-type N-terminal peptide that had a valine residue at 374, which corresponds to the methionine of the CYP2D6-met variant, was the same as that for human liver microsomes. We further confirmed that CYP2D6-met and CYP2D6-val expressed in human lymphoblastoid cells, both of which have methionine and valine, respectively, at position 374 and a human-type N-terminal peptide, exhibited the same enantioselectivities as those obtained from CYP2D6-met and CYP2D6-val expressed in the Hep G2 cell system. These results indicate that the amino acid at 374 of CYP2D6 is one of the key factors influencing the enantioselectivity of BTL 4-hydroxylation.",
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AU - Narimatsu, Shizuo

AU - Kato, Rika

AU - Horie, Toshiharu

AU - Ono, Satoshi

AU - Tsutsui, Michio

AU - Yabusaki, Yoshiyasu

AU - Ohmori, Shigeru

AU - Kitada, Mitsukazu

AU - Ichioka, Takao

AU - Shimada, Noriaki

AU - Kato, Ryuichi

AU - Ishikawa, Tsutomu

PY - 1999

Y1 - 1999

N2 - The enantioselectivity of 4-hydroxylation of bunitrolol (BTL), a β- adrenoceptor blocking drug, was studied in microsomes from human liver, human hepatoma (Hep G2) cells expressing CYP2D6, and lymphoblastoid cells expressing CYP2D6. Kinetics in human liver microsomes showed that the Vmax value for (+)-BTL was 2.1-fold that of (-)-BTL, and that the Km value for (+)-BTL was lower than that for the (-)antipode, resulting in the intrinsic clearance (Vmax/Km) of (+)-BTL being 2.1-fold over its (-)-antipode. CYP2D6 (CYP2D6-met) expressed in Hep G2 cells had a methionine residue at position 373 of the amino acid sequence and a rat-type N-terminal peptide (MELLNGTGLWSM) instead of the human-type (MGLEALVPLAVIV), and showed enantioselectivity of [(+)-BTL <(-)-BTL] for the rate of BTL 4- hydroxylation. In contrast, enantioselectivity [(+)-BTL > (-)-BTL] for Hep G2-CYP2D6 (CYP2D6-val) with a human-type N-terminal peptide that had a valine residue at 374, which corresponds to the methionine of the CYP2D6-met variant, was the same as that for human liver microsomes. We further confirmed that CYP2D6-met and CYP2D6-val expressed in human lymphoblastoid cells, both of which have methionine and valine, respectively, at position 374 and a human-type N-terminal peptide, exhibited the same enantioselectivities as those obtained from CYP2D6-met and CYP2D6-val expressed in the Hep G2 cell system. These results indicate that the amino acid at 374 of CYP2D6 is one of the key factors influencing the enantioselectivity of BTL 4-hydroxylation.

AB - The enantioselectivity of 4-hydroxylation of bunitrolol (BTL), a β- adrenoceptor blocking drug, was studied in microsomes from human liver, human hepatoma (Hep G2) cells expressing CYP2D6, and lymphoblastoid cells expressing CYP2D6. Kinetics in human liver microsomes showed that the Vmax value for (+)-BTL was 2.1-fold that of (-)-BTL, and that the Km value for (+)-BTL was lower than that for the (-)antipode, resulting in the intrinsic clearance (Vmax/Km) of (+)-BTL being 2.1-fold over its (-)-antipode. CYP2D6 (CYP2D6-met) expressed in Hep G2 cells had a methionine residue at position 373 of the amino acid sequence and a rat-type N-terminal peptide (MELLNGTGLWSM) instead of the human-type (MGLEALVPLAVIV), and showed enantioselectivity of [(+)-BTL <(-)-BTL] for the rate of BTL 4- hydroxylation. In contrast, enantioselectivity [(+)-BTL > (-)-BTL] for Hep G2-CYP2D6 (CYP2D6-val) with a human-type N-terminal peptide that had a valine residue at 374, which corresponds to the methionine of the CYP2D6-met variant, was the same as that for human liver microsomes. We further confirmed that CYP2D6-met and CYP2D6-val expressed in human lymphoblastoid cells, both of which have methionine and valine, respectively, at position 374 and a human-type N-terminal peptide, exhibited the same enantioselectivities as those obtained from CYP2D6-met and CYP2D6-val expressed in the Hep G2 cell system. These results indicate that the amino acid at 374 of CYP2D6 is one of the key factors influencing the enantioselectivity of BTL 4-hydroxylation.

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KW - Bunitrolol 4-hydroxylation

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KW - Enantioselectivity

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