Ispitivanje procesiranja scw4p proteina stanične stijenke u stanicama kex2 i 5ypsdeltakex2 mutanata kvasca S. Cerevisiae

undergraduate thesis

• Title (english): Investigation of the processing of yeast cell wall protein scw4p in the kex2 and 5ypsdeltakex2 mutant of S. Cerevisiae
• Author: Martina Glavan
• Mentor: Renata Teparić (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Chemistry and Biochemistry); (Laboratory for Biochemistry); Zagreb
• Defense date and country: 2015-07-08, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Protein Scw4p jedan je od najzastupljenijih nekovalentno vezanih proteina u staničnoj stijenci kvasca, međutim njegova uloga još uvijek nije otkrivena. Dosadašnjim istraživanjima pokazano je da se Scw4p protein može vezati u staničnu stijenku putem dva različita mehanizma - nekovalentno i kovalentno. Pretpostavlja se da se ovisno o procesiranju Scw4p pojavljuje u tri forme koje se razlikuju po molekulskoj masi te da način vezanja ovisi o procesiranju Scw4p pomoću proteaze Kex2p i/ili japsinskih proteaza, pri čemu se procesirani oblici proteina vežu nekovalentnim interakcijama dok se neprocesirani oblik Scw4p veže pretežno kovalentnim interakcijama u staničnu stijenku. Cilj ovog rada bio je Western blot analizom Scw4p izoliranog iz 5ypsΔkex2 mutanta kvasca te kex2 mutanta kvasca transformiranog nativnim i mutiranim oblicima gena SCW4 dobiti bolji uvid u procesiranje Scw4p proteina. U sklopu ovog rada ispitivan je i utjecaj prekomjerne sinteze Scw4p proteina na sposobnost preživljavanja stanica divljeg tipa, kex2, 5ypsΔ i 5ypsΔkex2 mutanata kako bi se prikupile informacije o mogućoj fiziološkoj ulozi i eventualnim razlikama u aktivnosti različitih formi ovog proteina u stijenci.

Abstract (english)

Protein Scw4p is one of the most abundant non-covalently linked cell wall proteins in yeast and its role in the cell has not yet been discovered. It was previously shown that the protein is linked to the yeast cell wall by two different mechanisms - covalently and non-covalently. It is assumed that proteolytic enzymes which might have a role in processing of Scw4p are Kex2p and a family of aspartic proteases called yapsins and that non-covalently bound forms of Scw4p underwent the proteolytic processing, while the covalently bound Scw4p was not processed. Depending on the processing three forms of Scw4p in the cell wall with different molecular weights could be found. In this work Western blot analysis of Scw4p protein, isolated from 5ypsΔkex2 and kex2 mutant yeast transformed with native and mutated forms of the gene SCW4, was performed in order to get a better insight into the processing of Scw4p protein. Another object of this study was to determinate the impact of protein Scw4p overexpression on the viability of wild-type cells, kex2, 5ypsΔ and 5ypsΔkex2 mutants to gather information about the possible physiological role and possible differences in the activities of the various forms of this protein in the yeast cell wall.

• Keywords: Saccharomyces cerevisiae; stanična stijenka kvasca; protein Scw4p; Kex2p; japsinske proteaze
• Keywords (english): Saccharomyces cerevisiae; yeast cell wall; Scw4p protein; Kex2p protease; aspartic proteases yapsins
• Language: croatian
• URN:NBN: urn:nbn:hr:159:357315
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-04-25 12:21:26