title: A new genetic defect in human CYP2C19: mutation of the initiation codon is responsible for poor metabolism of S-mephenytoin.
abstract: The 4'-hydroxylation of the S-enantiomer of the anticonvulsant drug mephenytoin exhibits a genetic polymorphism in humans. This polymorphism shows marked interracial heterogeneity, with the poor metabolizer (PM) phenotype representing 2 to 5% of Caucasian and 13 to 23% of Asian populations. Two defective CYP2C19 alleles, CYP2C19*2 and CYP2C19*3, have been described which account for approximately 87% of Caucasian and > 99% of Oriental PM alleles. The present study identifies a new allele (CYP2C19*4) in Caucasian PMs which contains an A-->G mutation in the initiation codon. A new polymerase chain reaction-restriction fragment length polymorphism genotyping test was developed, and the incidence of this allele was examined in a European Caucasian population which had been phenotyped for mephenytoin metabolism. One of nine putative PMs was heterozygous for CYP2C19*2/CYP2C19*4, which suggests that CYP2C19*4 represents a defective allele. Six of the seven remaining putative PMs available for genotyping were explained by CYP2C19*2. The frequency of the CYP2C19*4 allele in Caucasians was 0.6%. An additional Caucasian PM from a separate study was also heterozygous for CYP2C19*2 and CYP2C19*4. To verify that CYP2C19*4 represented a defective CYP2C19 allele, the initiation codon of the normal CYP2C19*1 cDNA was mutated to a GTG, and both cDNAs were expressed in yeast. Recombinant CYP2C19 protein was detected by Western blot analysis of colonies transformed with CYP2C19*1 cDNA, but not in those transformed with CYP2C19*4 cDNA. The two cDNAs were also used in an in vitro coupled transcription/translation assay. CYP2C19 protein was translated only from the CYP2C19*1 allele. These data indicate that CYP2C19*4 represents a new PM allele.
authors.affiliation: National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina, USA.
authors.forename: R J
authors.initials: RJ
authors.lastname: Ferguson
authors.forename: S M
authors.initials: SM
authors.lastname: De Morais
authors.forename: S
authors.initials: S
authors.lastname: Benhamou
authors.forename: C
authors.initials: C
authors.lastname: Bouchardy
authors.forename: J
authors.initials: J
authors.lastname: Blaisdell
authors.forename: G
authors.initials: G
authors.lastname: Ibeanu
authors.forename: G R
authors.initials: GR
authors.lastname: Wilkinson
authors.forename: T C
authors.initials: TC
authors.lastname: Sarich
authors.forename: J M
authors.initials: JM
authors.lastname: Wright
authors.forename: P
authors.initials: P
authors.lastname: Dayer
authors.forename: J A
authors.initials: JA
authors.lastname: Goldstein
mesh_terms: D000483:Alleles; D000927:Anticonvulsants / Q000378:metabolism*; D001189:Aryl Hydrocarbon Hydroxylases; D003062:Codon; D065731:Cytochrome P-450 CYP2C19; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D005260:Female; D006801:Humans; D008297:Male; D008617:Mephenytoin / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000235:genetics*; D009154:Mutation
chemical_list: D000927:Anticonvulsants; D003062:Codon; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19; D008617:Mephenytoin