Uno Yasuhiro, Kawabata Izumi, Ushirozako Genki, Tsukiyama-Kohara Kyoko, Ishizuka Mayumi, Mizukawa Hazuki, Murayama Norie, Yamazaki Hiroshi
Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima-city, Kagoshima 890-0065, Japan.
Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima-city, Kagoshima 890-0065, Japan.
Biochem Pharmacol. 2025 Jun;236:116894. doi: 10.1016/j.bcp.2025.116894. Epub 2025 Mar 26.
Pigs are often used in drug metabolism studies because of their evolutionary proximity to humans, including similarities in their cytochromes P450 (P450s or CYPs). In the current study, the following cDNAs of novel CYP4Fs were isolated and characterized: dog CYP4F22 and CYP4F140; cat CYP4F22 and CYP4F140; pig CYP4F22, CYP4F52, CYP4F53, CYP4F54, CYP4F56, and CYP4F176; and tree shrew CYP4F22. Previously identified pig CYP4F55 cDNA was also isolated. These CYP4F cDNAs contained open reading frames of 522-531 amino acids and shared high sequence identities (60-92 %) with human CYP4Fs. Dog CYP4F3a and CYP4F3b cDNAs were also identified but lacked the 3' end of the coding region. Phylogenetic analysis of amino acid sequences showed that these CYP4Fs were clustered in a species-dependent manner, except for CYP4F3, CYP4F22, and CYP4F140, which were clustered in an isoform-dependent manner. All CYP4F genes, containing 12 coding exons, formed a gene cluster at the corresponding location of the genome in each species. Among the tissue samples analyzed, dog and cat CYP4F140 mRNAs were more abundantly expressed in liver/testis and kidney, respectively. Preferential expression of pig CYP4F mRNAs were found in liver, small intestine, and/or kidney, where the most abundant were CYP4F56, CYP4F52, and CYP4F176 mRNAs, respectively. Enzyme assays using recombinant proteins revealed that all these CYP4Fs oxidized the human CYP4F substrate arachidonic acid at the ω-position, indicating that they are functional enzymes. These findings suggest that dog, cat, pig, and tree shrew CYP4Fs have similar functional characteristics to human CYP4Fs.
由于猪在进化上与人类接近，包括其细胞色素P450（P450或CYP）的相似性，猪常用于药物代谢研究。在本研究中，分离并鉴定了以下新型CYP4F的cDNA：犬CYP4F22和CYP4F140；猫CYP4F22和CYP4F140；猪CYP4F22、CYP4F52、CYP4F53、CYP4F54、CYP4F56和CYP4F176；以及树鼩CYP4F22。还分离了先前鉴定的猪CYP4F55 cDNA。这些CYP4F cDNA包含522 - 531个氨基酸的开放阅读框，与人CYP4F具有高度的序列同一性（60 - 92%）。犬CYP4F3a和CYP4F3b cDNA也被鉴定出来，但缺少编码区的3'末端。氨基酸序列的系统发育分析表明，除了以同工型依赖方式聚类的CYP4F3、CYP4F22和CYP4F140外，这些CYP4F以物种依赖的方式聚类。所有包含12个编码外显子的CYP4F基因在每个物种基因组的相应位置形成一个基因簇。在所分析的组织样本中，犬和猫的CYP4F140 mRNA分别在肝脏/睾丸和肾脏中表达更丰富。在猪的肝脏、小肠和/或肾脏中发现了CYP4F mRNA的优先表达，其中最丰富的分别是CYP4F56、CYP4F52和CYP4F176 mRNA。使用重组蛋白的酶分析表明，所有这些CYP4F在ω位氧化人CYP4F底物花生四烯酸，表明它们是功能性酶。这些发现表明，犬、猫、猪和树鼩的CYP4F具有与人类CYP4F相似的功能特征。
