title: Functional characterization of a common CYP4F11 genetic variant and identification of functionally defective CYP4F11 variants in erythromycin metabolism and 20-HETE synthesis.
abstract: CYP4F11, together with CYP4F2, plays an important role in the synthesis of 20-hydroxyeicosatetraenoic acid (20-HETE) from arachidonic acid. We identified 21 variants by whole exome sequencing, including 4 non-synonymous variants in Korean subjects. The proteins of the wild-type CYP4F11 and the four coding variants (C276R, D315N, D374Y, and D446N) were expressed in Escherichia coli DH5α cells and purified to give cytochrome P450-specific carbon monoxide difference spectra. Wild-type CYP4F2 was also expressed and purified to compare its activity with the CYP4F11 wild-type. Wild-type CYP4F11 exhibited the highest maximal clearance for erythromycin N-demethylase activity followed by the variants D374Y, D446N, C276R, and D315N. In particular, the CYP4F11 D315N protein showed about 50% decrease in intrinsic clearance compared to the wild type. The ability of wild-type CYP4F11 and the variants to synthesize 20-HETE from arachidonic acid was similar; the CYP4F11 D315N variant, however, showed only 68% of wild-type activity. Furthermore, the ability of CYP4F2 to synthesize 20-HETE was 1.7-fold greater than that of CYP4F11. Overall, our results suggest that the metabolism of CYP4F11 substrates may be reduced in individuals carrying the CYP4F11 D315N genetic variant and individuals carrying the common D446N CYP4F11 variant likely exhibit comparable 20-HETE synthesis as individuals expressing wild-type CYP4F11.
authors.affiliation: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea.
authors.forename: Myeongjin
authors.initials: M
authors.lastname: Yi
authors.affiliation: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea.
authors.forename: Sun-Ah
authors.initials: SA
authors.lastname: Cho
authors.affiliation: Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709, USA.
authors.forename: Jungki
authors.initials: J
authors.lastname: Min
authors.affiliation: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea.
authors.forename: Dong Hyun
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea; Department of Clinical Pharmacology, Inje University Busan Paik Hospital, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea. Electronic address: phshinjg@gmail.com.
authors.forename: Jae-Gook
authors.initials: JG
authors.lastname: Shin
authors.affiliation: Department of Pharmacology and Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Bokji-ro 75, Busanjin-gu, Busan 47392, South Korea. Electronic address: 2sujun@inje.ac.kr.
authors.forename: Su-Jun
authors.initials: SJ
authors.lastname: Lee
mesh_terms: D019943:Amino Acid Substitution; D016718:Arachidonic Acid / Q000737:chemistry / Q000378:metabolism; D000072470:Cytochrome P450 Family 4 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004917:Erythromycin / Q000737:chemistry; D059472:Exome; D005260:Female; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis / Q000737:chemistry; D008297:Male; D020125:Mutation, Missense; D011994:Recombinant Proteins
chemical_list: D006893:Hydroxyeicosatetraenoic Acids; D011994:Recombinant Proteins; D016718:Arachidonic Acid; D004917:Erythromycin; C055987:20-hydroxy-5,8,11,14-eicosatetraenoic acid; D000072470:Cytochrome P450 Family 4; C492722:CYP4F11 protein, human
keywords: 20-Hydroxyeicosatetraenoic acid; Arachidonic acid; CYP4F11; CYP4F2; Erythromycin; Genetic variants