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

72 Scopus citations

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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Qu, W., Bradbury, J. A., Tsao, C. C., Maronpot, R., Harry, G. J., Parker, C. E., Davis, L. S., Breyer, M. D., Waalkes, M. P., Falck, J. R., Chen, J., Rosenberg, R. L., & 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