Cytochrome P450 CYP2J9, a New Mouse Arachidonic Acid ω-1 Hydroxylase Predominantly Expressed in Brain

Wei Qu, J. Alyce Bradbury, Cheng Chung Tsao, Robert Maronpot, G. Jean Harry, Carol E. Parker, Linda S. Davis, Matthew D. Breyer, Michael P. Waalkes, J R Falck, Jianyong Chen, Robert L. Rosenberg, Darryl C. Zeldin

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

A cDNA encoding a new cytochrome P450 was isolated from a mouse brain library. Sequence analysis reveals that this 1,958-base pair cDNA encodes a 57-58-kDa 502-amino acid polypeptide that is 70-91% identical to CYP2J subfamily P450s and is designated CYP2J9. Recombinant CYP2J9 was co-expressed with NADPH-cytochrome P450 oxidoreductase (CYPOR) in Sf9 cells using a baculovirus system. Microsomes of CYP2J9/CYPOR-transfected cells metabolize arachidonic acid to 19-hydroxyeicosatetraenoic acid (HETE) thus CYP2J9 is enzymologically distinct from other P450s. Northern analysis reveals that CYP2J9 transcripts are present at high levels in mouse brain. Mouse brain microsomes biosynthesize 19-HETE. RNA polymerase chain reaction analysis demonstrates that CYP2J9 mRNAs are widely distributed in brain and most abundant in the cerebellum. Immunoblotting using an antibody raised against human CYP2J2 that cross-reacts with CYP2J9 detects a 56-kDa protein band that is expressed in cerebellum and other brain segments and is regulated during postnatal development. In situ hybridization of mouse brain sections with a CYP2J9-specific riboprobe and immunohistochemical staining with the anti-human CYP2J2 IgG reveals abundant CYP2J9 mRNA and protein in cerebellar Purkinje cells. Importantly, 19-HETE inhibits the activity of recombinant P/Q-type Ca2+ channels that are known to be expressed preferentially in cerebellar Purkinje cells and are involved in triggering neurotransmitter release. Based on these data, we conclude that CYP2J9 is a developmentally regulated P450 that is abundant in brain, localized to cerebellar Purkinje cells, and active in the biosynthesis of 19-HETE, an eicosanoid that inhibits activity of P/Q-type Ca2+ channels. We postulate that CYP2J9 arachidonic acid products play important functional roles in the brain.

Original languageEnglish (US)
Pages (from-to)25467-25479
Number of pages13
JournalJournal of Biological Chemistry
Volume276
Issue number27
DOIs
StatePublished - Jul 6 2001

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Mixed Function Oxygenases
Arachidonic Acid
Cytochrome P-450 Enzyme System
Brain
Hydroxyeicosatetraenoic Acids
Purkinje Cells
Microsomes
Cerebellum
cytochrome P-450 CYP2J9 (murine)
Complementary DNA
Sf9 Cells
NADPH-Ferrihemoprotein Reductase
Messenger RNA
Eicosanoids
Baculoviridae
Polymerase chain reaction
Biosynthesis
DNA-Directed RNA Polymerases
Immunoblotting
Base Pairing

ASJC Scopus subject areas

  • Biochemistry

Cite this

Qu, W., Bradbury, J. A., Tsao, C. C., Maronpot, R., Harry, G. J., Parker, C. E., ... Zeldin, D. C. (2001). Cytochrome P450 CYP2J9, a New Mouse Arachidonic Acid ω-1 Hydroxylase Predominantly Expressed in Brain. Journal of Biological Chemistry, 276(27), 25467-25479. https://doi.org/10.1074/jbc.M100545200

Cytochrome P450 CYP2J9, a New Mouse Arachidonic Acid ω-1 Hydroxylase Predominantly Expressed in Brain. / Qu, Wei; Bradbury, J. Alyce; Tsao, Cheng Chung; Maronpot, Robert; Harry, G. Jean; Parker, Carol E.; Davis, Linda S.; Breyer, Matthew D.; Waalkes, Michael P.; Falck, J R; Chen, Jianyong; Rosenberg, Robert L.; Zeldin, Darryl C.

In: Journal of Biological Chemistry, Vol. 276, No. 27, 06.07.2001, p. 25467-25479.

Research output: Contribution to journalArticle

Qu, W, Bradbury, JA, Tsao, CC, Maronpot, R, Harry, GJ, Parker, CE, Davis, LS, Breyer, MD, Waalkes, MP, Falck, JR, Chen, J, Rosenberg, RL & Zeldin, DC 2001, 'Cytochrome P450 CYP2J9, a New Mouse Arachidonic Acid ω-1 Hydroxylase Predominantly Expressed in Brain', Journal of Biological Chemistry, vol. 276, no. 27, pp. 25467-25479. https://doi.org/10.1074/jbc.M100545200
Qu, Wei ; Bradbury, J. Alyce ; Tsao, Cheng Chung ; Maronpot, Robert ; Harry, G. Jean ; Parker, Carol E. ; Davis, Linda S. ; Breyer, Matthew D. ; Waalkes, Michael P. ; Falck, J R ; Chen, Jianyong ; Rosenberg, Robert L. ; Zeldin, Darryl C. / Cytochrome P450 CYP2J9, a New Mouse Arachidonic Acid ω-1 Hydroxylase Predominantly Expressed in Brain. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 27. pp. 25467-25479.
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abstract = "A cDNA encoding a new cytochrome P450 was isolated from a mouse brain library. Sequence analysis reveals that this 1,958-base pair cDNA encodes a 57-58-kDa 502-amino acid polypeptide that is 70-91{\%} identical to CYP2J subfamily P450s and is designated CYP2J9. Recombinant CYP2J9 was co-expressed with NADPH-cytochrome P450 oxidoreductase (CYPOR) in Sf9 cells using a baculovirus system. Microsomes of CYP2J9/CYPOR-transfected cells metabolize arachidonic acid to 19-hydroxyeicosatetraenoic acid (HETE) thus CYP2J9 is enzymologically distinct from other P450s. Northern analysis reveals that CYP2J9 transcripts are present at high levels in mouse brain. Mouse brain microsomes biosynthesize 19-HETE. RNA polymerase chain reaction analysis demonstrates that CYP2J9 mRNAs are widely distributed in brain and most abundant in the cerebellum. Immunoblotting using an antibody raised against human CYP2J2 that cross-reacts with CYP2J9 detects a 56-kDa protein band that is expressed in cerebellum and other brain segments and is regulated during postnatal development. In situ hybridization of mouse brain sections with a CYP2J9-specific riboprobe and immunohistochemical staining with the anti-human CYP2J2 IgG reveals abundant CYP2J9 mRNA and protein in cerebellar Purkinje cells. Importantly, 19-HETE inhibits the activity of recombinant P/Q-type Ca2+ channels that are known to be expressed preferentially in cerebellar Purkinje cells and are involved in triggering neurotransmitter release. Based on these data, we conclude that CYP2J9 is a developmentally regulated P450 that is abundant in brain, localized to cerebellar Purkinje cells, and active in the biosynthesis of 19-HETE, an eicosanoid that inhibits activity of P/Q-type Ca2+ channels. We postulate that CYP2J9 arachidonic acid products play important functional roles in the brain.",
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AU - Tsao, Cheng Chung

AU - Maronpot, Robert

AU - Harry, G. Jean

AU - Parker, Carol E.

AU - Davis, Linda S.

AU - Breyer, Matthew D.

AU - Waalkes, Michael P.

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