Abstract
Cytochrome P450 4F2 (CYP4F2) is an enzyme that is involved in the metabolism of arachidonic acid (AA), vitamin E and K, and xenobiotics including drugs. CYP4F2*3 polymorphism (rs2108622; c.1297G>A; p.Val433Met) has been associated with hypertension, ischemic stroke, and variation in the effectiveness of the anticoagulant drug warfarin. In this study, we characterized wild-type CYP4F2 and 28 CYP4F2 variants, including a Val433Met substitution, detected in 8380 Japanese subjects. The CYP4F2 variants were heterologously expressed in 293FT cells to measure the concentrations of CYP4F2 variant holoenzymes using carbon monoxide–reduced difference spectroscopy, where the wild type and 18 holoenzyme variants showed a peak at 450 nm. Kinetic parameters [Vmax, substrate concentration producing half of Vmax (S50), and intrinsic clearance (CLint) as Vmax/S50] of AA ω-hydroxylation were determined for the wild type and 21 variants with enzyme activity. Compared with the wild type, two variants showed significantly decreased CLint values for AA ω-hydroxylation. The values for seven variants could not be determined because no enzymatic activity was detected at the highest substrate concentration used. Three-dimensional structural modeling was performed to determine the reason for reduced enzymatic activity of the CYP4F2 variants. Our findings contribute to a better understanding of CYP4F2 variant-associated diseases and possible future therapeutic strategies.
SIGNIFICANCE STATEMENT CYP4F2 is involved in the metabolism of arachidonic acid and vitamin K, and CYP4F2*3 polymorphisms have been associated with hypertension and variation in the effectiveness of the anticoagulant drug warfarin. This study presents a functional analysis of 28 CYP4F2 variants identified in Japanese subjects, demonstrating that seven gene polymorphisms cause loss of CYP4F2 function, and proposes structural changes that lead to altered function.
Footnotes
- Received May 8, 2023.
- Accepted September 25, 2023.
M.H. was supported by grants from the Japan Agency for Medical Research and Development (AMED) [Grant JP19kk0305009] and The Naito Foundation. S.S., S.T., and K.K. were supported by grants from AMED [Grants JP17km0105001 and JP21tm0124005]. This research was also supported in part by the Tohoku Medical Megabank Project: Promoting Public Utilization of Advanced Research Infrastructure and the Sharing and Administrative Network for Research Equipment, funded by the Ministry of Education, Culture, Sports, Science, and Technology (MEXT). S.T. and K.K. were supported by grants for the Facilitation of R&D Platform for AMED Genome Medicine Support from AMED [Grant JP20km0405001]. Y.S. was supported by the Japan Society for the Promotion of Science [Grant 21K15291]. This research was supported by Research Support Project for Life Science and Drug Discovery [Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)] from AMED [Grant JP22ama121019].
The authors declare no conflict of interest.
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- Copyright © 2023 by The American Society for Pharmacology and Experimental Therapeutics
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