Cloning, Sequencing, and Tissue Expression of CYP3A27, a New Member of the CYP3A Subfamily from Embryonic and Adult Rainbow Trout Livers

Lee, Su-Jun; Wang-Buhler, Jun-Lan; Çok, İsmet; Yu, Tinghe; Yang, Yea-Huey; Miranda, Cristobal L.; Lech, John J.; Buhler, Donald R.

doi:10.1006/abbi.1998.0943

Cited by 50 publications

(26 citation statements)

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“…Amino acid comparisons and immunological studies between trout CYP3A27 and human CYP3A4 were reported (Miranda et al, 1991;Lee et al, 1998) and the present observation of nifedipine metabolism could further support a similar structure function relationship between human CYP3A4 and trout CYP3A27. CYP3A27 is the first designated CYP3A form in the aquatic species (Lee et al, 1998) and the precise functional study requires cDNA directed expression of this gene to remove other background cytochrome P450s from the trout liver microsomes. In this study, we report the heterologous expression and the functional study of CYP3A27 gene by using recombinant baculovirus system.…”

Section: Lee and Buhlersupporting

confidence: 80%

“…The study of trout CYP3A form was initiated from the purification of LMC5 P450, which showed catalytic activity against testosterone and progesterone (Miranda et al, 1989) and cross-reacted with human CYP3A4 antibodies (Miranda et al, 1991). Previous research suggested that there is more than one CYP3A form in trout (Lee et al, 1998). It is possible, therefore, that the purification of specific CYP3A27 from the trout liver did not separate completely this isoform from the other CYP3A forms.…”

Section: Discussionmentioning

confidence: 99%

“…In a previous study, we reported the cloning of a new CYP3A form, CYP3A27, from rainbow trout (Lee et al, 1998). CYP3A27 was obtained by cDNA library screening using the antibodies generated against purified trout cytochrome P450 LMC5, which displayed significant activity against steroids and reacted with the antibodies of human CYP3A4 (Miranda et al, 1989(Miranda et al, , 1991.…”

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Functional Properties of a Rainbow Trout CYP3A27 Expressed by Recombinant Baculovirus in Insect Cells

Lee

Buhler²

2002

Drug Metab Dispos

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ABSTRACT:Cytochrome P450 3A27 (CYP3A27) is highly expressed in liver and intestine of rainbow trout (Oncorhynchus mykiss). In many animal species, the intestine and liver are responsible for the first-pass metabolism of a wide range of xenobiotics. To help determine its physiological role, the catalytic capabilities of CYP3A27 protein were examined. An open reading frame of CYP3A27 in pFastBac donor plasmid was transferred to the baculovirus genome (bacmid DNA) through Tn7 site-specific transposition in DH10Bac competent cells. The CYP3A27 cDNA was positioned under the control of the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus. The recombinant baculovirus containing a fulllength CYP3A27 cDNA (Bv-3A27) was then transfected into Spodoptera frugiperda (Sf9) insect cells for overexpression of CYP3A27 protein. The expressed CYP3A27 protein (714 pmol/mg total protein) exhibited a maximum CO-reduced spectrum at 450 nm at 72 h postinfection after addition of 1 g/ml exogenous hemin. The expressed CYP3A27 protein comigrated with the purified trout LMC5 cytochrome P450 (P450) and was recognized by anti-P450 LMC5 IgG on Western blot analysis. The expressed CYP3A27 protein was reconstituted with human NADPH-cytochrome P450 reductase and cytochrome b 5 . The reconstitution system showed catalytic activities for the 6␤-, 2␤-, and 16␤-hydroxylation of testosterone at 1.428, 0.043, 0.034 nmol/min/nmol CYP3A27, respectively, and the dehydrogenation of nifedipine at 50 pmol/min/nmol CYP3A27. The present results demonstrated that the baculovirus system is useful for the production of the functional aquatic CYP3A form and that CYP3A27 has the capability to metabolize steroid hormone as reported for mammalian CYP3A forms.

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Section: Lee and Buhlersupporting

confidence: 80%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Functional Properties of a Rainbow Trout CYP3A27 Expressed by Recombinant Baculovirus in Insect Cells

Lee

Buhler²

2002

Drug Metab Dispos

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“…Some vertebrate CYP3As also are widely expressed, for example in liver, brain, kidney, testis, gonad, stomach, and heart of rainbow trout (Lee et al, 1998). The Ciona genes also are expressed at different stages of development indicating a potential stage-specific functional role.…”

Section: Discussionmentioning

confidence: 99%

Isolation and phylogeny of novel cytochrome P450 genes from tunicates (Ciona spp.): A CYP3 line in early deuterostomes?

Verslycke

Goldstone

Stegeman

2006

Molecular Phylogenetics and Evolution

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Cytochromes P450 (CYPs) form a gene superfamily involved in the biotransformation of numerous endogenous and exogenous natural and synthetic compounds. In humans, CYP3A4 is regarded as one of the most important CYPs due to its abundance in liver and its capacity to metabolize more than 50% of all clinically used drugs. It has been suggested that all CYP3s arose from a common ancestral gene lineage that diverged between 800 and 1100 million years ago, before the deuterostome-protostome split. While CYP3s are well known in mammals and have been described in lower vertebrates, they have not been reported in non-vertebrate deuterostomes. Members of the genus Ciona belong to the tunicates, whose lineage is thought to be the most basal among the chordates, and from which the vertebrate line diverged. Here we describe the cloning, exon-intron structure, phylogeny, and estimated expression of four novel genes from Ciona intestinalis. We also describe the gene structure and phylogeny of homologous genes in Ciona savignyi. Comparing these genes with other members of the CYP clan 3, show that the Ciona sequences bear remarkable similarity to vertebrate CYP3A genes, and may be an early deuterostome CYP3 line.

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“…These sex differences in metabolism are now known to reflect sex differences in the expression of specific, individual hepatic P450 enzymes and other DMEs (Shapiro et al, 1995). Sex-linked differences in P450-dependent drug and steroid metabolism are also found in hamster (Teixeira and Gil, 1991;Sakuma et al, 1994), dog (Lin et al, 1996;Hay Kraus et al, 2000), chicken (Pampori and Shapiro, 1993), and fish (Gray et al, 1991;Lee et al, 1998). Sex differences in human hepatic P450-catalyzed drug metabolism are well documented, but are less dramatic than in the rat (Tanaka, 1999;Parkinson et al, 2004;Scandlyn et al, 2008).…”

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Sex Differences in the Expression of Hepatic Drug Metabolizing Enzymes

Waxman

Holloway²

2009

Mol Pharmacol

634

584

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Sex differences in pharmacokinetics and pharmacodynamics characterize many drugs and contribute to individual differences in drug efficacy and toxicity. Sex-based differences in drug metabolism are the primary cause of sex-dependent pharmacokinetics and reflect underlying sex differences in the expression of hepatic enzymes active in the metabolism of drugs, steroids, fatty acids and environmental chemicals, including cytochromes P450 (P450s), sulfotransferases, glutathione transferases, and UDP-glucuronosyltransferases. Studies in the rat and mouse liver models have identified more than 1000 genes whose expression is sex-dependent; together, these genes impart substantial sexual dimorphism to liver metabolic function and pathophysiology. Sex differences in drug metabolism and pharmacokinetics also occur in humans and are due in part to the female-predominant expression of CYP3A4, the most important P450 catalyst of drug metabolism in human liver. The sexually dimorphic expression of P450s and other liver-expressed genes is regulated by the temporal pattern of plasma growth hormone (GH) release by the pituitary gland, which shows significant sex differences. These differences are most pronounced in rats and mice, where plasma GH profiles are highly pulsatile (intermittent) in male animals versus more frequent (nearly continuous) in female animals. This review discusses key features of the cell signaling and molecular regulatory mechanisms by which these sex-dependent plasma GH patterns impart sex specificity to the liver. Moreover, the essential role proposed for the GH-activated transcription factor signal transducer and activator of transcription (STAT) 5b, and for hepatic nuclear factor (HNF) 4␣, as mediators of the sexdependent effects of GH on the liver, is evaluated. Together, these studies of the cellular, molecular, and gene regulatory mechanisms that underlie sex-based differences in liver gene expression have provided novel insights into the physiological regulation of both xenobiotic and endobiotic metabolism.

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Cloning, Sequencing, and Tissue Expression of CYP3A27, a New Member of the CYP3A Subfamily from Embryonic and Adult Rainbow Trout Livers

Cited by 50 publications

References 32 publications

Functional Properties of a Rainbow Trout CYP3A27 Expressed by Recombinant Baculovirus in Insect Cells

Functional Properties of a Rainbow Trout CYP3A27 Expressed by Recombinant Baculovirus in Insect Cells

Isolation and phylogeny of novel cytochrome P450 genes from tunicates (Ciona spp.): A CYP3 line in early deuterostomes?

Sex Differences in the Expression of Hepatic Drug Metabolizing Enzymes

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