Determining the type of interaction between zebrafish Oct1 and endo- and xenobiotics (CROSBI ID 637211)
Prilog sa skupa u zborniku | sažetak izlaganja sa skupa
Podaci o odgovornosti
Mihaljević, Ivan ; Popović, Marta ; Žaja, Roko ; Maraković, Nikola ; Smital, Tvrtko
engleski
Determining the type of interaction between zebrafish Oct1 and endo- and xenobiotics
Organic cation transporters (OCT) are polyspecific membrane transporters from Solute Carrier superfamily (SLC), present in all vertebrates, where they play crucial role in homeostasis of organic cations, which encompass various endo- and xenobiotics. There are two Oct members in zebrafish, in respect to three members present from reptiles to human. Oct1 in zebrafish is dominantly expressed in kidney and liver, which indicates its potential compensatory role of human OCT1 and OCT2, which are dominantly expressed in liver and kidney, respectively. In our previous research we developed in vitro assay for functional characterization of Oct1, based on heterologous expression system in HEK293T cells. Using fluorescent substrates, we identified various interactors ranging from physiological compounds such as steroid hormones to the environmental contaminants such as organotin compounds and various pharmaceuticals. In present study, we tried to determine the type of interaction with previously identified interactors using classic Michaelis-Menten kinetics. However, results revealed mixed type of interaction with several compounds, pointing to more complex active region of Oct1. In order to characterize Oct1 active region in more detail, we performed homology modeling and molecular docking and identified potentially crucial amino acid residues. These amino acids are also conserved within active region of human OCT1 and 2, which further confirms their crucial role. We determined that Lys215, Trp218, Arg440 and Ser470 have important role as donors of hydrogen atom in H-bond, whereas Phe160, Ser163, Leu164, Thr443, Gln447 and Cys550 contribute to formation of H-bond as acceptors of hydrogen atom. Amino acid residues Phe160, Tyr170, Trp219, Trp355, Arg440, and Phe447 were shown to form aromatic bonds between Oct1 and tested compounds. Our further research will focus on determination of roles these amino acid residues play within active region using site-directed mutagenesis.
Slc22 ; Oct1 ; zebrafish ; functional characterization ; homology modeling
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Podaci o prilogu
64-64.
2016.
objavljeno
Podaci o matičnoj publikaciji
Abstract Book - International FishMed Conference on Zebrafish Research
Bialek-Wyrzykowska, Urszula
Varšava: International Institute of Molecular and Cell Biology in Warsaw
Podaci o skupu
International FishMed Conference on Zebrafish Research
poster
18.03.2016-19.03.2016
Varšava, Poljska