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Functional characterization of rainbow trout (Oncorhynchus mykiss) Abcg2a (Bcrp) transporter

Žaja, Roko; Popović, Marta; Lončar, Jovica; Smital, Tvrtko (2016) Functional characterization of rainbow trout (Oncorhynchus mykiss) Abcg2a (Bcrp) transporter. Comparative Biochemistry and Physiology - Part C: Comparative Pharmacology, 190 . pp. 15-23. ISSN 0306-4492

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ABCG2 (BCRP – breast cancer resistance protein) belongs to the ATP-binding cassette (ABC) superfamily. It plays an important role in the disposition and elimination of xeno- and endobiotics and/or their metabolites in mammals. Likewise, the protective role of ABC transporters, including Abcg2, has been reported for aquatic organisms. In our previous study we have cloned the full gene sequence of rainbowtrout (Oncorhynchusmykiss) Abcg2a and showed its high expression in liver and primary hepatocytes. Based on those insights, the main goal of this study was to perform a detailed functional characterization of trout Abcg2a using insect ovary cells (Spodoptera frugiperda, Sf9) as a heterologous expression system. Membrane vesicles preparations from Sf9 cells were used for the ATPase assay determinations and basic biochemical properties offishAbcg2a versushumanABCG2 have been compared. A series of 39 physiologically and/or environmentally relevant substances was then tested on interaction with trout Abcg2a and human ABCG2. Correlation analysis reveals highly similar pattern of activation and inhibition. Significant activation of trout Abcg2a ATPase was observed for prazosin, doxorubicine, sildenafil, furosemid, propranolol, fenofibrate and pheophorbide. Pesticides showed either a weak activation (malathione) or strong (endosulfan) to weak (chlorpyrifos, fenoxycarb, DDE) inhibition of trout Abcg2a ATPase while the highest activation was obtained for benzo(a)pyrene, curcumine and testosterone. In conclusion, data fromthis study offer the first characterization of fish Abcg2a, reveal potent interactors among physiologically or environmentally relevant substances and point to similarities regarding strengths and interactor preferences between human ABCG2 and fish Abcg2a.

Item Type: Article
Uncontrolled Keywords: ABCG2/BCRP ; Breast cancer resistance protein ; Rainbow trout ; Insect ovary cells (Spodoptera frugiperda ; Sf9) ; ATPase determinations ; Substrate/interactor preferences
Subjects: NATURAL SCIENCES > Biology
NATURAL SCIENCES > Biology > Biochemistry and Molecular Biology
NATURAL SCIENCES > Biology > Genetics, Evolution and Phylogenetics
Divisions: Division for Marine and Enviromental Research
Project titleProject leaderProject codeProject type
Ekotoksikološko značenje ABC transportnih proteina u vodenih organizama-Tvrtko Smital098-0982934-2745MZOS
Depositing User: Marta Popović
Date Deposited: 07 Nov 2018 16:28
DOI: 10.1016/j.cbpc.2016.07.005

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