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

Roko Zaja1, Marta Popović1, Jovica Lončar1

  • 1Laboratory for Molecular Ecotoxicology, Division for Marine and Environmental Research, Ruđer Bošković Institute, Zagreb, Croatia.

Comparative Biochemistry and Physiology. Toxicology & Pharmacology : CBP
|August 1, 2016
PubMed

Insights

This study functionally characterized rainbow trout Abcg2a, revealing its interactions with various substances. Fish Abcg2a shares similarities with human ABCG2 in substrate preferences, aiding in understanding xenobiotic transport.

Area of Science:

  • Environmental Toxicology
  • Biochemistry
  • Molecular Biology

Background:

  • ATP-binding cassette (ABC) transporters, like ABCG2, are crucial for xenobiotic and endobiotic disposition in mammals.
  • The protective role of ABC transporters, including Abcg2, has been documented in aquatic organisms.
  • Rainbow trout (Oncorhynchus mykiss) Abcg2a was previously cloned and shown to be highly expressed in liver and hepatocytes.

Purpose of the Study:

  • To perform a detailed functional characterization of rainbow trout Abcg2a.
  • To compare the biochemical properties of fish Abcg2a with human ABCG2.
  • To identify substances that interact with trout Abcg2a.

Main Methods:

  • Heterologous expression of trout Abcg2a in Spodoptera frugiperda (Sf9) insect ovary cells.
  • ATPase assay determinations using membrane vesicles from Sf9 cells.
  • Testing the interaction of 39 physiologically and/or environmentally relevant substances with trout Abcg2a and human ABCG2.

Main Results:

  • Trout Abcg2a ATPase activity was significantly activated by prazosin, doxorubicin, sildenafil, furosemide, propranolol, fenofibrate, and pheophorbide.
  • Pesticides showed varied effects, with endosulfan strongly inhibiting trout Abcg2a ATPase, while malathion weakly activated it.
  • Benzo(a)pyrene, curcumin, and testosterone showed the highest activation of trout Abcg2a ATPase, with a highly similar pattern of activation and inhibition observed between fish Abcg2a and human ABCG2.

Conclusions:

  • This study provides the first functional characterization of fish Abcg2a.
  • Potent interactors among physiologically and environmentally relevant substances were identified for trout Abcg2a.
  • Similarities in interactor preferences and strengths between human ABCG2 and fish Abcg2a were highlighted, suggesting conserved functions.
Keywords:
ABCG2/BCRPATPase determinationsBreast cancer resistance proteinInsect ovary cells (Spodoptera frugiperda, Sf9)Rainbow troutSubstrate/interactor preferences

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