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Molecular cloning and functional characterization of a rainbow trout liver Oatp
Author(s) -
Steiner Konstanze,
Dietrich Daniel,
Hagenbuch Bruno
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1047.10
Subject(s) - rainbow trout , complementary dna , biology , open reading frame , biochemistry , amino acid , chemistry , gene , microbiology and biotechnology , peptide sequence , fishery , fish <actinopterygii>
Microcystins (MCs) are natural occurring toxins produced by cyanobacteria in aquatic ecosystems. MCs are known hepatotoxicants in fish and mammals, whereby toxicity in mammals has been demonstrated to result from Oatp‐mediated MC‐uptake into hepatocytes. To investigate whether MC‐uptake in fish liver is Oatp‐mediated, the goal of this study was to clone and functionally characterize a rainbow trout liver Oatp. We prepared a cDNA library from rainbow trout liver mRNA and isolated a full length Oatp. To functionally characterize this Oatp we transiently transfected HEK293 cells and determined uptake of radiolabelled Oatp substrates. The isolated cDNA consists of 2773 bp and has an open reading frame of 2115 bp, encoding a 705‐amino acid protein. Phylogentic analysis suggests that the isolated protein is Oatp1d1. Functional characterization revealed that Oatp1d1 can mediate the uptake of various Oatp substrates including bromosulfophthalein (BSP), D‐penicillamine(2,5)enkephalin (DPDPE), estradiol‐17β‐glucuronide (E17βG), estrone‐3‐sulfate, taurocholate, and methotrexate. In conclusion, we have isolated and characterized a multispecific Oatp1d1 from rainbow trout liver that may be instrumental in MC uptake.