Simple confirmation methods for rare but impaired variants of human flavin-containing monooxygenase 3 (FMO3) found in an updated genome resource databank

被引:1
作者
Shimizu, Makiko [1 ]
Makiguchi, Miaki [1 ]
Yokota, Yuka [1 ]
Shimamura, Erika [1 ]
Matsuta, Moegi [1 ]
Nakamura, Yuria [1 ]
Harano, Mizuki [1 ]
Yamazaki, Hiroshi [1 ,2 ]
机构
[1] Showa Pharmaceut Univ, Lab Drug Metab & Pharmacokinet, Machida, Tokyo 1948543, Japan
[2] Showa Pharmaceut Univ, Lab Drug Metab & Pharmacokinet, 3-3165 Higashi Tamagawa Gakuen, Machida, Tokyo 1948543, Japan
基金
日本学术振兴会;
关键词
FMO3; Polymerase chain reaction; Restriction fragment length polymorphism; TRIMETHYLAMINURIA; GENE; POLYMORPHISMS;
D O I
10.1016/j.dmpk.2023.100528
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Forty-seven new nonsense or missense human flavin-containing monooxygenase 3 (FMO3) variants were recently identified in an updated Japanese population reference panel. Of these, 20 rare single-nucleotide substitutions resulted in moderately or severely impaired FMO3 activity. To easily identify these 20 FMO3 variants (2 stop codon mutations, 2 frameshifts, and 16 amino-acid substitutions) in the clinical setting, simple confirmation methods for impaired FMO3 variants are proposed using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) or allele-specific PCR methods. Using PCR-RFLP, FMO3 variants p.Arg51Gly, p. Met66Lys, p.Asn80Lys, p.Val151Glu, p.Val187fsTer25, p.Gly193Arg, p.Val283Ala, p.Asp286His, p.Val382Ala, and p.Phe451Leu were digested by the designated restriction enzymes and confirmed using reference cDNAs. In contrast, the FMO3 variants p.Gly39Val, p.Arg238Ter, p.Arg387Cys, p.Arg387His, p.Leu457Trp, and p. Met497Arg were not digested, whereas the wild type was digested. FMO3 variants p.Gly11Asp, p.Lys416fsTer72, p.Gln427Ter, and p.Thr453Pro were confirmed using allele-specific PCR systems. The previously identified FMO3 p.Arg500Ter variant has a relatively high frequency and was differentiated from p.Arg500Gln in two steps, i.e., enzyme restriction followed by allele-specific PCR, similar to the method for p.Arg387Cys and p.Arg387His. These systems should facilitate easy detection in the clinical setting of FMO3 variants in Japanese subjects susceptible to low drug clearance possibly caused by impaired FMO3 function.
引用
收藏
页数:7
相关论文
共 24 条
[21]   A novel mutation in the flavin-containing monooxygenase 3 gene, FMO3, that causes fish-odour syndrome:: activity of the mutant enzyme assessed by proton NMR spectroscopy [J].
Murphy, HC ;
Dolphin, CT ;
Janmohamed, A ;
Holmes, HC ;
Michelakakis, H ;
Shephard, EA ;
Chalmers, RA ;
Phillips, IR ;
Iles, RA .
PHARMACOGENETICS, 2000, 10 (05) :439-451
[22]   Novel variants and haplotypes of human flavin-containing monooxygenase 3 gene associated with Japanese subjects suffering from trimethylaminuria [J].
Shimizu, Makiko ;
Yoda, Hiromi ;
Igarashi, Narumi ;
Makino, Miki ;
Tokuyama, Emi ;
Yamazaki, Hiroshi .
XENOBIOTICA, 2019, 49 (10) :1244-1250
[23]   Oxidative stress-responsive transcription factor NRF2 is not indispensable for the human hepatic Flavin-containing monooxygenase-3 (FMO3) gene expression in HepG2 cells [J].
Rudraiah, Swetha ;
Gu, Xinsheng ;
Hines, Ronald N. ;
Manautou, Jose E. .
TOXICOLOGY IN VITRO, 2016, 31 :54-59
[24]   Complex mechanism underlying transcriptional control of the haplotyped flavin-containing monooxygenase 3 (FMO3) gene in Japanese:: Different regulation between mutations in 5′-upstream distal region and common element in proximal region [J].
Shimizu, Makiko ;
Murayama, Norie ;
Nagashima, Satomi ;
Fujieda, Masaki ;
Yamazaki, Hiroshi .
DRUG METABOLISM AND PHARMACOKINETICS, 2008, 23 (01) :54-58