Recombinant Human Cytochrome P450 4X1 (CYP4X1)
Description
Expression and Purification of CYP4X1
CYP4X1 has been expressed in Escherichia coli using codon-optimized cDNA constructs to enhance its expression levels. The highest expression levels were achieved with a bicistronic construct that included the cDNA for human NADPH-P450 reductase, reaching up to 300-450 nmol P450/L culture . This recombinant protein has been shown to oxidize anandamide, an important endocannabinoid, to 14,15-EET ethanolamide, suggesting a potential role in neurovascular signaling pathways .
Biological Functions and Significance
CYP4X1 is primarily expressed in brain regions and skin, as indicated by quantitative PCR analysis . Its role in the brain suggests involvement in neurological functions, possibly through the metabolism of endocannabinoids like anandamide. Additionally, CYP4X1 has been implicated in the progression of colorectal cancer (CRC), where its overexpression is associated with metastasis, poor prognosis, and shorter survival times .
Role in Colorectal Cancer
Recent studies have demonstrated that CYP4X1 expression is significantly higher in CRC tissues compared to normal colon tissues. High CYP4X1 expression correlates with advanced TNM stages, poor tumor differentiation, deeper invasion, and lymph node metastasis . Downregulation of CYP4X1 in CRC cells inhibits cell proliferation, invasion, migration, and colony formation, suggesting its potential as a therapeutic target .
Table 1: CYP4X1 Expression in Colorectal Cancer
Table 2: CYP4X1 Expression Levels in Different Tissues
| Tissue Type | CYP4X1 Expression Level |
|---|---|
| Brain Regions | High |
| Skin | High |
| Normal Colon Tissue | Low or absent |
| Colorectal Cancer Tissue | Significantly higher than normal tissue |
References
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Kim, S., et al. (2023). Expression of CYP4X1 in colorectal carcinoma is associated with metastasis and poor prognosis. Research Square.
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Stark, K., et al. (2006). Expression and purification of orphan cytochrome P450 4X1 and oxidation of anandamide. PMC.
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[Anonymous] (2025). CYP4X1 Expression Is Associated with Metastasis and Poor Prognosis in Colorectal Cancer. PMC.
Product Specs
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Target Background
Recombinant Human Cytochrome P450 4X1 (CYP4X1) is a monooxygenase that selectively catalyzes the epoxidation of the final double bond in the arachidonoyl moiety of anandamide, potentially influencing endocannabinoid signaling. It lacks hydroxylase activity toward various fatty acids, steroids, and prostaglandins. Mechanistically, it utilizes molecular oxygen, incorporating one oxygen atom into the substrate and reducing the second to water. The required two electrons are provided by NADPH via cytochrome P450 reductase (CPR).
- Key residues crucial for arachidonic acid and anandamide binding to human CYP4X1 have been identified. PMID: 25595103
- PPARα plays a role in CYP4X1 regulation, while glucocorticoid and progesterone receptors are involved in CYP4Z1 gene activation. PMID: 15797250
