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master's thesis
Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju
diplomski rad
Split: University of Split, Faculty of Chemistry and Technology, 2016. urn:nbn:hr:167:121316
Ledić, Antonio
University of Split
Faculty of Chemistry and Technology
Division of Chemistry
University of Split
School of Medicine

Institutional repository: Repository of the Faculty of chemistry and technology

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Ledić, A. (2016). Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad (Master's thesis). Split: University of Split, Faculty of Chemistry and Technology; Split: University of Split, School of Medicine. Retrieved from https://urn.nsk.hr/urn:nbn:hr:167:121316

Ledić, Antonio. "Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad." Master's thesis, University of Split, Faculty of Chemistry and Technology; University of Split, School of Medicine, 2016. https://urn.nsk.hr/urn:nbn:hr:167:121316

Ledić, Antonio. "Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad." Master's thesis, University of Split, Faculty of Chemistry and Technology; University of Split, School of Medicine, 2016. https://urn.nsk.hr/urn:nbn:hr:167:121316

Ledić, A. (2016). 'Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad', Master's thesis, University of Split, Faculty of Chemistry and Technology; University of Split, School of Medicine, accessed 24 November 2024, https://urn.nsk.hr/urn:nbn:hr:167:121316

Ledić A. Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad [Master's thesis]. Split: University of Split, Faculty of Chemistry and Technology; Split: University of Split, School of Medicine; 2016 [cited 2024 November 24] Available at: https://urn.nsk.hr/urn:nbn:hr:167:121316

A. Ledić, "Utjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad", Master's thesis, University of Split, Faculty of Chemistry and Technology, Split; University of Split, School of Medicine, Split, 2016. Available at: https://urn.nsk.hr/urn:nbn:hr:167:121316

Metadata
TitleUtjecaj dimerizacije NIX/BNIP3L receptura na mitofagiju : diplomski rad
Title (english)The role of NIX/BNIP3L receptor on mitophagy : master's thesis
AuthorAntonio Ledić
MentorIvana Novak (mentor)
Committee memberIvana Novak (predsjednik povjerenstva)
GranterUniversity of Split
Faculty of Chemistry and Technology
(Division of Chemistry)
Split
GranterUniversity of Split
School of Medicine
Split
Defense date and country2016-10-12, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Pharmacy
Abstract
Mitofagija je važan proces kojim stanica uklanja oštećene mitohondrije. Protein Nix ima važnu ulogu u mitofagiji kao receptor te se pretpostavlja da se aktivira dimerizacijom. Cilj ovoga istraživanja bio je utvrditi da li fosforilacija serinskog ostataka S212 na C- terminalnom kraju mitohondrijskog proteina Nix utječe na njegovu dimerizaciju, a samim time i njegovu funkciju. U tu svrhu upotrijebljeni su konstrukti EGFP-NixWT, EGFP- NixS212A i EGFP-NixS212E. GST pull-down metodom određena je interakcija EGFP-Nix proteinskih mutanata s autofagijskim proteinom LC3A. Imunofluorescencijskom mikroskopijom promatrana je kolokalizacija mutanata proteina Nix i proteina vanjske mitohondrijske membrane Tom20 kako bi se utvrdio pravilan smještaj proteina Nix u vanjskoj mitohondrijskoj membrani. Imunofluorescencijska mikroskopija upotrebljena je i za promatranje kolokalizacije autofagosomalnog proteina LC3A i mutanata proteina Nix. Rezultati su pokazali da mutacije proteina Nix koristene u eksperimentu ne utjecu na interakciju sa autofagosomalniim proteinom LC3A te ju svi uspjesno ostvaruju ali postoji razlika u mogucnosti stvaraja dimera. Mutanta S212E ppokazuje nemogucnost stvaranja dimera. Imunofluorescencijskom mikroskopijom dokazano je da su mutante proteina Nix lokalizirane na vanjskoj mitohondrijskoj membrani što ukazuje da promijena navedenog aminokiselinskog slijeda ne mijenja njegovu lokalizaciju na mitohondriju. Za potvrdu uloge fosforiliranog serina S212 te njegov utjecaj na dimerizaciju i funkciju Nix proteina potrebni su dodatni eksperimenti.
Abstract (english)
Cell uses mitophagy to eliminate damaged mitochondria. Nix protein has a key role in mitophagy as receptor and it is proposed that it is activated by dimeryzation. .The objective of this study was to determine how phosphorylation of serine residue, S212, on the C- terminal end of Nix mitochondrial protein affects his dimerizationa and therefore its function as a mitophagy receptor. EGFP-NixWT, EGFP-NixS212A and EGFP - NixS212E constructs were used. The interaction of EGFP- Nix protein mutants with autophagosomal protein LC3A was determinated using GST pull-down method. Colocalization of Nix mutants with outer mitochondrial membrane protein Tom20 was observed using immunofluorescent microscopy. Immunofluorescent microscopy was also used to observe colocalization of autophagosomal protein LC3A and Nix mutants. Results showed that all Nix mutants used in the study interact with autophagosomal protein LC3A eventhough the dimerisation of NixS212E is abolished. Further, immunofluorescent microscopy analysis showed that Nix mutants are present on outer mitochondrion membrane which confirms that changes od S212 aminoacid does not influence its localization to mitochondria..Tu further confirm if phosphorylation of serine S212 influences Nix dimerisation and therefore, its function, further experiments are essential.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:167:121316
Study programmeTitle: Pharmacy
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2016-11-24 12:06:46