Razvoj sustava za ispitivanje interakcija Mate3 proteina zebrice s okolišnim tvarima

master's thesis

• Title (english): Development of an assay for studying interactions of zebrafish Mate3 protein with environmental chemicals
• Author: Lana Mišić
• Mentor: Igor Stuparević (mentor)
• Committee member: Igor Slivac (član povjerenstva)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2021-09-24, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Ubrzani rast populacije i tehnološki napredak rezultiraju sve većom izloženošću organizama okolišnim ksenobioticima. Nakon početne apsorpcije i distribucije u organizmu, slijedi detoksikacija i izlučivanje ksenobiotika iz organizma, najčešće putem urina ili žuči. U procesu izlučivanja, važnu ulogu imaju MATE sekundarni transporteri, prisutni u svim carstvima živog svijeta, koji kataliziraju elektroneutralan antiport protona i organskih kationa. Uspješno smo, pomoću Flp-In rekombinantnog sustava, razvili stabilnu HEK293T staničnu liniju sa prekomjerno eksprimiranim Mate3 transportnim proteinom zebrice (Danio rerio) koja se smatra jednim od najvažnijih modelnih organizama u ekotoksikološkim studijama. Uspješno smo optimirali visoko procesivan transportni sustav za ispitivanje interakcija Mate3 proteina s okolišnim tvarima, što će omogućiti detaljnu karakterizaciju uloge Mate3 proteina u detoksikacijskim procesima zebrice.

Abstract (english)

Rapid growth of populations and technological advancement has resulted in growing exposure of organisms to the innumerable environmental xenobiotics. After the initial absorption and distribution throughout the organism, xenobiotics are detoxified and excreted outside of the organism, mainly by urine and bile. In the excretion process, the secondary MATE transporters, found in all kingdoms of life, bear a significant role in electroneutral H+/organic cation antiport. By using Flp-In recombinant system, we have successfully developed a stable HEK293T cell line with overexpressed Mate3 transport protein found in zebrafish (Danio rerio), that is considered one of the most important model organisms in ecotoxicological studies. Also, we have successfully optimized highly processive transport assay for identifying interactions between Mate3 protein and environmental substances, which will allow detailed characterization of the Mate3 role in detoxification processes in zebrafish.

• Keywords: zebrica; ksenobiotici; Mate; transport; optimizacija
• Keywords (english): zebrafish; xenobiotics; Mate; transport; optimization
• Language: croatian
• URN:NBN: urn:nbn:hr:159:570741
• Study programme: Title: Molecular Biotechnology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka molekularne biotehnologije (magistar/magistra inženjer/inženjerka molekularne biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-10-22 09:28:37