Rat CYP2D2, not 2D1, is functionally conserved with human CYP2D6 in endogenous morphine formation

Nadja Grobe; Toni M. Kutchan; Meinhart H. Zenk

doi:10.1016/j.febslet.2012.05.021

Rat CYP2D2, not 2D1, is functionally conserved with human CYP2D6 in endogenous morphine formation

Nadja Grobe, Toni M. Kutchan, Meinhart H. Zenk

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

The assumption that CYP2D1 is the corresponding rat cytochrome to human CYP2D6 has been revisited using recombinant proteins in direct enzyme assays. CYP2D1 and 2D2 were incubated with known CYP2D6 substrates, the three morphine precursors thebaine, codeine and (R)-reticuline. Mass spectrometric analysis showed that rat CYP2D2, not 2D1, catalyzed the 3-O-demethylation reaction of thebaine and codeine. In addition, CYP2D2 incubated with (R)-reticuline generated four products corytuberine, pallidine, salutaridine and isoboldine while rat CYP2D1 was completely inactive. This intramolecular phenol-coupling reaction follows the same mechanism as observed for CYP2D6. Michaelis-Menten kinetic parameters revealed high catalytic efficiencies for rat CYP2D2. These findings suggest a critical evaluation of other commonly accepted, however untested, CYP2D1 substrates.

Original language	English
Pages (from-to)	1749-1753
Number of pages	5
Journal	FEBS Letters
Volume	586
Issue number	13
DOIs	https://doi.org/10.1016/j.febslet.2012.05.021
State	Published - Jun 21 2012

Keywords

CYP2D1 and CYP2D2
CYP2D6
LC-MS/MS analysis
Mammalian morphine biosynthesis

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10.1016/j.febslet.2012.05.021

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title = "Rat CYP2D2, not 2D1, is functionally conserved with human CYP2D6 in endogenous morphine formation",

abstract = "The assumption that CYP2D1 is the corresponding rat cytochrome to human CYP2D6 has been revisited using recombinant proteins in direct enzyme assays. CYP2D1 and 2D2 were incubated with known CYP2D6 substrates, the three morphine precursors thebaine, codeine and (R)-reticuline. Mass spectrometric analysis showed that rat CYP2D2, not 2D1, catalyzed the 3-O-demethylation reaction of thebaine and codeine. In addition, CYP2D2 incubated with (R)-reticuline generated four products corytuberine, pallidine, salutaridine and isoboldine while rat CYP2D1 was completely inactive. This intramolecular phenol-coupling reaction follows the same mechanism as observed for CYP2D6. Michaelis-Menten kinetic parameters revealed high catalytic efficiencies for rat CYP2D2. These findings suggest a critical evaluation of other commonly accepted, however untested, CYP2D1 substrates.",

keywords = "CYP2D1 and CYP2D2, CYP2D6, LC-MS/MS analysis, Mammalian morphine biosynthesis",

author = "Nadja Grobe and Kutchan, {Toni M.} and Zenk, {Meinhart H.}",

note = "Funding Information: The authors would like to thank Dr. Leslie Hicks and Dr. Sophie Alvarez, Donald Danforth Plant Science Center, for sequencing the commercially available CYP2D1 and CYP2D2. We would like to thank Megan Rolf for excellent technical assistance. This work was supported by the National Institutes of Health Grant # 5R21DA024418 and the Mallinckrodt Foundation, St. Louis .",

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AU - Grobe, Nadja

AU - Kutchan, Toni M.

AU - Zenk, Meinhart H.

N1 - Funding Information: The authors would like to thank Dr. Leslie Hicks and Dr. Sophie Alvarez, Donald Danforth Plant Science Center, for sequencing the commercially available CYP2D1 and CYP2D2. We would like to thank Megan Rolf for excellent technical assistance. This work was supported by the National Institutes of Health Grant # 5R21DA024418 and the Mallinckrodt Foundation, St. Louis .

PY - 2012/6/21

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N2 - The assumption that CYP2D1 is the corresponding rat cytochrome to human CYP2D6 has been revisited using recombinant proteins in direct enzyme assays. CYP2D1 and 2D2 were incubated with known CYP2D6 substrates, the three morphine precursors thebaine, codeine and (R)-reticuline. Mass spectrometric analysis showed that rat CYP2D2, not 2D1, catalyzed the 3-O-demethylation reaction of thebaine and codeine. In addition, CYP2D2 incubated with (R)-reticuline generated four products corytuberine, pallidine, salutaridine and isoboldine while rat CYP2D1 was completely inactive. This intramolecular phenol-coupling reaction follows the same mechanism as observed for CYP2D6. Michaelis-Menten kinetic parameters revealed high catalytic efficiencies for rat CYP2D2. These findings suggest a critical evaluation of other commonly accepted, however untested, CYP2D1 substrates.

AB - The assumption that CYP2D1 is the corresponding rat cytochrome to human CYP2D6 has been revisited using recombinant proteins in direct enzyme assays. CYP2D1 and 2D2 were incubated with known CYP2D6 substrates, the three morphine precursors thebaine, codeine and (R)-reticuline. Mass spectrometric analysis showed that rat CYP2D2, not 2D1, catalyzed the 3-O-demethylation reaction of thebaine and codeine. In addition, CYP2D2 incubated with (R)-reticuline generated four products corytuberine, pallidine, salutaridine and isoboldine while rat CYP2D1 was completely inactive. This intramolecular phenol-coupling reaction follows the same mechanism as observed for CYP2D6. Michaelis-Menten kinetic parameters revealed high catalytic efficiencies for rat CYP2D2. These findings suggest a critical evaluation of other commonly accepted, however untested, CYP2D1 substrates.

KW - CYP2D1 and CYP2D2

KW - CYP2D6

KW - LC-MS/MS analysis

KW - Mammalian morphine biosynthesis

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ER -

Rat CYP2D2, not 2D1, is functionally conserved with human CYP2D6 in endogenous morphine formation

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