title: Human CYP4F12 genetic polymorphism: identification and functional characterization of seven variant allozymes.
abstract: The human cytochrome CYP4F12 has been shown to be metabolically active toward inflammatory mediators and exogenous compounds such as antihistaminic drugs. We recently identified a genetic polymorphism within the promoter region, associated with a decreased level of enzyme expression. In the present study, we report the further identification of single nucleotide polymorphisms in the coding sequence of the CYP4F12 gene. A polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis of DNA samples from 53 unrelated French Caucasians, allowed the identification of ten mutations, comprising seven missense mutations, 31C>T (Leu11Phe), 38C>T (Pro13Leu), 47C>T (Met16Thr), 4759G>A (Asp76Asn), 4801G>A (Val90Leu), 8896C>T (Arg188Cys) and 23545G>A (Gly522Ser). Their functional impact toward ebastine hydroxylation was evaluated using heterologous expression in Saccharomyces cerevisiae cells of site-directed mutated cDNA variants. Five out seven variants did not exhibit any significant difference in CYP4F12 catalytic activity, whereas two variants, Val90Ile and Arg188Cys, displayed significant changes in their Michaelis-Menten (Km, Vm) parameters. These data on CYP4F12 genetic polymorphism provide tools for further studies of association with pathological processes involving an inflammatory component and with variations in anti-histaminic drug response.
authors.affiliation: Equipe d'accueil 2679, Faculté de Médecine de Lille, Pôle Recherche, Lille, France. christelle.cauffiez@caramail.com
authors.forename: Christelle
authors.initials: C
authors.lastname: Cauffiez
authors.forename: Florian
authors.initials: F
authors.lastname: Klinzig
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Rat
authors.forename: Gilles
authors.initials: G
authors.lastname: Tournel
authors.forename: Delphine
authors.initials: D
authors.lastname: Allorge
authors.forename: Dany
authors.initials: D
authors.lastname: Chevalier
authors.forename: Nicolas
authors.initials: N
authors.lastname: Pottier
authors.forename: Tonio
authors.initials: T
authors.lastname: Lovecchio
authors.forename: Jean-Frédéric
authors.initials: JF
authors.lastname: Colombel
authors.forename: Michel
authors.initials: M
authors.lastname: Lhermitte
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Lo-Guidice
authors.forename: Franck
authors.initials: F
authors.lastname: Broly
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D002384:Catalysis; D004252:DNA Mutational Analysis; D015870:Gene Expression; D006801:Humans; D006899:Mixed Function Oxygenases / Q000235:genetics*; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C422180:CYP4F12 protein, human