title: Functional analysis of novel polymorphisms in the human SLCO1A2 gene that encodes the transporter OATP1A2.
abstract: The solute carrier organic anion transporting polypeptide 1A2 (OATP1A2, SLCO1A2) is implicated in the cellular influx of a number of drugs. We identified five novel single nucleotide polymorphisms (SNPs) in coding exons of the SLCO1A2 gene in a cohort of subjects: G550A, G553A, G673A, A775C, and G862A, that encoded the OATP1A2 variants E184K, D185N, V255I, T259P, and D288N, respectively. The function and expression of these variant transporters were assessed in HEK-293 cells. We found that the novel variants, E184K, D185N, T259P, and D288N, were associated with impaired estrone-3-sulfate, imatinib, and methotrexate transport (∼20-50% of wild-type control); function was retained by OATP1A2-V255I. From biotinylation assays, the decreased function of these variants was due, at least in part, to impaired plasma membrane expression. The four loss-of-function variants were studied further using mutagenesis to produce variants that encode residues with different charges or steric properties. From immunoblotting, the replacement of negatively charged residues at amino acid positions 184 and 185 impaired membrane expression, while either a positive or negative charge at residue 288 supported the correct membrane targeting of OATP1A2. Replacement of T259 with bulky residues disrupted transporter stability. From molecular models, E184, D185, and D288 were located near several charged residues such that intramolecular ionic interactions may stabilize the transporter structure. Individuals who carry these novel SNPs in the SLCO1A2 gene may be at risk from impaired efficacy or enhanced toxicity during treatment with drugs that are substrates for OATP1A2.
authors.affiliation: Faculty of Pharmacy, The University of Sydney, Sydney, NSW, 2006, Australia, fanfan.zhou@sydney.edu.au.
authors.forename: Fanfan
authors.initials: F
authors.lastname: Zhou
authors.forename: Jian
authors.initials: J
authors.lastname: Zheng
authors.forename: Ling
authors.initials: L
authors.lastname: Zhu
authors.forename: Andreas
authors.initials: A
authors.lastname: Jodal
authors.forename: Pei H
authors.initials: PH
authors.lastname: Cui
authors.forename: Mark
authors.initials: M
authors.lastname: Wong
authors.forename: Howard
authors.initials: H
authors.lastname: Gurney
authors.forename: W Bret
authors.initials: WB
authors.lastname: Church
authors.forename: Michael
authors.initials: M
authors.lastname: Murray
mesh_terms: D005815:Genetic Code / Q000502:physiology; D014644:Genetic Variation / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D027361:Organic Anion Transporters / Q000737:chemistry* / Q000502:physiology*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*
chemical_list: D027361:Organic Anion Transporters; C512988:SLCO1A2 protein, human
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