Biochemical and Biophysical Research Communications
Identification of a novel cytochrome P450, CYP4X1, with unique localization specific to the brain☆,☆☆
Section snippets
Materials and methods
Sequence analysis. Data on human CYP4X1 ESTs were obtained from the UniGene database [21]. The BLAST algorithm was used for searches of the HTGS sections in the GenBank [20]. Multiple alignments were prepared by Jotun Hein method of the Megaline program (DNA Star, Madison, WI). The genomic sequence used for analysis of the mouse cyp4x1 gene was a 234-kb long draft sequence (Accession No. AJ297131) [19] and the genomic sequences used for analysis of the human CYP4X1 gene were two 178-kb long
CYP4X1 cDNA
RT-PCR using primers designed toward two putative exons of a mouse cyp4x1 genomic sequence yielded a partial CYP4X1 cDNA sequence from rat brain, whereas no PCR bands were generated from either rat liver, kidney or cultured astrocytes. A 3′-RACE method in combination with the usage of a 5′-UTR specific primer for the mouse gene was used to obtain the sequence of the full coding region of the rat CYP4X1 cDNA. The nucleotide sequence contained an open reading frame of 1521-bp (Fig. 1). The 3′
Discussion
In this study, we report the cDNA cloning and initial characterization of a rat brain specific P450 isoform belonging to a novel CYP4 subfamily. The deduced amino acid sequence of the enzyme exhibits 41–51% identity with that of members of the rat CYP4 subfamilies 4A, 4B, and 4F. According to the P450 nomenclature standard, this enzyme did not fulfill the criteria for belonging to any of the characterized mammalian subfamilies [1]. The enzyme was designated rat CYP4X1. It has been hypothesized
Acknowledgements
This work was supported by NIH/NINDS R01 NS32321-05, NIH/NHLBI P01 HL59996, VA Merit 3440-02P, and Wenner-Gren Foundations. We thank Dr. Pompon (CNRS, France) for the generous gift of the S. cerevisiae strain W(R).
References (28)
Cytochrome P450 and the individuality of species
Arch. Biochem. Biophys.
(1999)- et al.
Neurosteroids: biosynthesis and function of these novel neuromodulators
Front. Neuroendocrinol.
(2000) - et al.
Dopamine formation from tyramine by CYP2D6
Biochem. Biophys. Res. Commun.
(1998) - et al.
Microsomal cytochrome P450-mediated liver and brain anandamide metabolism
Biochem. Pharmacol.
(1995) - et al.
Delivery and turnover of plasma-derived essential PUFAs in mammalian brain
J. Lipid Res.
(2001) The cytochrome P450 4 (CYP4) family
Gen. Pharmacol.
(1997)- et al.
Identification of CYP4F8 in human seminal vesicles as a prominent 19-hydroxylase of prostaglandin endoperoxides
J. Biol. Chem.
(2000) - et al.
Alternative splicing determines the function of CYP4F3 by switching substrate specificity
J. Biol. Chem.
(2001) - et al.
Positional specificity of rabbit CYP4B1 for ω-hydroxylation of short-medium chain fatty acids and hydrocarbons
Biochem. Biophys. Res. Commun.
(1998) - et al.
A transgenic mouse expressing human CYP4B1 in the liver
Biochem. Biophys. Res. Commun.
(2001)
Enhanced in vivo monooxygenase activities of mammalian P450s in engineered yeast cells producing high levels of NADPH-P450 reductase and human cytochrome b5
Gene
Structural alignments of P450s and extrapolations to the unknown
Methods Enzymol.
Neurosteroid hydroxylase CYP7B: vivid reporter activity in dentate gyrus of gene-targeted mice and abolition of a widespread pathway of steroid and oxysterol hydroxylation
J. Biol. Chem.
Cytochrome P450 CYP2J9, a new mouse arachidonic acid ω-1 hydroxylase predominantly expressed in brain
J. Biol. Chem.
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2017, Pharmacology and TherapeuticsCitation Excerpt :Multiple members of the CYP4 family, which have hydroxylase activity, have been shown to be expressed in brain homogenates, brain sections, the basilar and middle cerebral arteries, cerebral microvessels, as well as in astrocytes, endothelial cells, vascular smooth muscle cells and neurons. All of these cells types therefore have the necessary machinery to produce 19- and 20-HETE (Al-Anizy et al., 2006; Alonso-Galicia, Hudetz, Shen, Harder, & Roman, 1999; Bylund, Zhang, & Harder, 2002; Carver et al., 2014; Dunn et al., 2008; Gebremedhin et al., 1998, 2000, 2016; Zhang, Falck, et al., 2017; Zhu et al., 2015). 20-HETE, but not 19-HETE have been detected in vascular smooth muscle cells, and more recently astrocytes and neurons (Gebremedhin, Yamaura, & Harder, 2008; Gebremedhin et al., 1998, 2016; Zhang, Falck, et al., 2017).
Expression of CYP4V2 in human THP1 macrophages and its transcriptional regulation by peroxisome proliferator-activated receptor gamma
2017, Toxicology and Applied PharmacologyExpression of CYP 4A ω-hydroxylase and formation of 20-hydroxyeicosatetreanoic acid (20-HETE) in cultured rat brain astrocytes
2016, Prostaglandins and Other Lipid MediatorsCitation Excerpt :They couple neuronal activity to local blood flow as a component of the neurovascular unit (NVU) [1,2,3]. A variety of membrane-derived lipids is emerging as important regulators of cerebrovascular function under physiological and pathological conditions [4–12]. Enzymes of the cytochrome P450 (CYP) gene family are expressed in different cell types in the brain and catalyze conversion of AA released by phospholipase A2 to a variety of fatty acid signaling molecules [5–7,12–16].
Phylogenetic and functional analyses of the cytochrome P450 family 4
2012, Molecular Phylogenetics and EvolutionCitation Excerpt :In our analysis and others (Savas et al., 2005), the highest CYP4X expression was found in the trachea (Fig. 4A) although the biological importance of this had not been determined. The expression of CYP4X in tissues associated with neuronal function, such as brain, was high (Bylund et al., 2002; Savas et al., 2005; Stark et al., 2008) and our data agrees. CYP expression in the brain is limited; brain CYP expression was only 0.5–2% of hepatic CYP expression (Hedlund et al., 2001).
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Abbreviations: CYP, cytochrome P450; P450, cytochrome P450; EST, expressed sequence tag; HTGS, high throughput genomic sequences; RACE, rapid amplification of cDNA ends; RT-PCR, reverse trancription-polymerase chain reaction; UTR, untranslated region.