Abstract
Pharmaceuticals and their metabolites constitute a class of xenobiotics commonly found in aquatic environments which may cause toxic effects in aquatic organisms. Several different lipophilic molecules, including some pharmaceuticals, can bind to fatty acid-binding proteins (FABPs), a group of evolutionarily related cytoplasmic proteins that belong to the intracellular lipid-binding protein (iLBP) family. An oyster FABP genome-wide investigation was not available until a recent study on gene organization, protein structure, and phylogeny of Crassostrea gigas iLBPs. Higher transcript levels of the C. gigas FABP2 gene were found after exposure to sewage and pharmaceuticals. Because of its relevance as a potential biomarker of aquatic contamination, in this study, recombinant FABP2 from C. gigas (CgFABP2) was successfully cloned, expressed, and purified, and in vitro and in silico assays were performed using lipids and pharmaceuticals. This is the first characterization of a protein from the iLBP family in C. gigas. Homology modeling and molecular docking were used to evaluate the binding affinities of natural ligands (palmitic, oleic, and arachidonic acids) and pharmaceuticals (ibuprofen, sodium diclofenac, and acetaminophen). Among the tested fatty acids, CgFABP2 showed preference for palmitic acid. The selected pharmaceuticals presented a biphasic-binding mode, suggesting a different binding affinity with a preference for diclofenac. Therefore, the approach using circular dichroism and in silico data might be useful for ligand-binding screening in an invertebrate model organism.
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A Correction to this paper has been published: https://doi.org/10.1007/s11356-021-13135-x
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Acknowledgements
The authors would like to thank the Laboratório Central de Biologia Molecular Estrutural (CEBIME/UFSC) for use in their structural biology facilities.
Funding
This research was funded by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq Grant # 406104/2013-1 – PVE Fellowship). Juliana F. Tisca, MSc. and Tomás B. Pessatti, MSc. are PhD fellows at the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Fabiola S. Soares and Karin dos Santos were fellows at the CNPq. Dr. Flávia L. Zacchi is a postdoctoral fellow at the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES). Dr. Afonso Celso Dias Bainy is a recipient of the CNPq productivity fellowship. M. J. Bebianno is a recipient of the CNPq PVE fellowship and also acknowledges the support of the Portuguese Science Foundation (FCT) through the grant UID/MAR/00350/2013 attributed to CIMA, at the University of Algarve.
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JFT conducted the experiments, analyzed the data, and wrote the manuscript. TBP performed the experiment and analyzed the data. FSS performed the experiment. KdS performed the in silico experiments, analyzed the data and helped with writing the manuscript. VAO performed the circular dichroism and analyzed the data. FLZ, MJAFB, ACDB, and GR analyzed the data, and contributed in the discussions, writing process, and proofreading of the manuscript. All authors read and approved the final version of the manuscript.
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Tisca, J.F., dos Santos, K., Pessati, T.B. et al. Characterization of a fatty acid-binding protein from the Pacific oyster (Crassostrea gigas): pharmaceutical and toxicological implications. Environ Sci Pollut Res 28, 27811–27822 (2021). https://doi.org/10.1007/s11356-021-12645-y
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DOI: https://doi.org/10.1007/s11356-021-12645-y