master's thesis
Utjecaj Fe3+ iona na mitohondrije kvasca Saccharomyces cerevisiale W303

Martina Barbarić (2016)
Sveučilište u Rijeci
Medicinski fakultet
Katedra za kemiju i biokemiju
Metadata
TitleUtjecaj Fe3+ iona na mitohondrije kvasca Saccharomyces cerevisiale W303
AuthorMartina Barbarić
Mentor(s)Gordana Čanadi Jurešić (thesis advisor)
Abstract
Kvasac je čovječanstvu poznat od davnina kroz proizvodnju kruha, piva i vina. Danas se zna da je njegov potencijal mnogo veći. Zbog poznatog genoma i jednostavnog uzgoja postaje vodeći modelni eukariotski organizam u različitim znanstvenim istraživanjima kao i izučavanju štetnih utjecaja okolišnih čimbenika kao što je željezo. Ioni željeza u niskim koncentracijama esencijalni su za rast stanice, međutim u višim koncentracijama postaju toksični, induciraju nastanak ROS-a te posljedično uzrokuju promjene na staničnoj i molekularnoj razini. Cilj rada bio je ispitati utjecaj željeza na proteom i lipidom mitohondrija u čiju je svrhu korišten kvasac Saccharomyces cerevisiae W303. Analize su provedene na kvascu uzgajanom u prisustvu željeza kao i na netretiranom kvascu kao kontrolnom uzorku. Proteini mitohondrija razdvojeni su metodom 2D elektroforeze, obojeni bojilom Sypro Ruby te vizualizirani koristeći UV transiluminator. Usporedba proteinskih profila tretiranog i netretiranog kvasca izvršena je uz pomoć programa Dymension from Syngene. U tretiranom uzorku detektiran je manji broj proteina, 8 proteina pojačalo je, a samo jedan smanjio izričaj u odnosu na kontrolu. Također, pod utjecajem željeza došlo je do sinteze 5 novih proteina. Koristeći računalni program ProteinPilot v 4.5 uspješno su identificirana dva proteina: podjedinica enzima KGDH uključenog u ciklus limunske kiseline i CPR6 enzim iz skupine ciklofilina. Promjene na razini lipidoma određene su na temelju masnokselinskog sastava ukupnih mitohondrijskih lipida koristeći plinsku kromatografiju. Glavne promjene zabilježene u tretiranom uzorku bile su porast zasićenosti masnih kiselina i produljenje acilnih lanaca. Također, analiza lipida pokazala je smanjenje masenog udjela lipida prema proteinima u mitohondrijima kvasca tretiranog željezom.
Keywordsyeast iron mitochondrion
Committee MembersMarin Tota (committee chairperson)
Branka Blagović (committee member)
Gordana Čanadi Jurešić (committee member)
GranterSveučilište u Rijeci
Medicinski fakultet
Lower level organizational unitsKatedra za kemiju i biokemiju
PlaceRijeka
StateCroatia
Scientific field, discipline, subdisciplineBIOMEDICINE AND HEALTHCARE
Basic Medical Sciences
Medical Biochemistry
Study programme typeuniversity
Study levelgraduate
Study programmeStudy of Sanitary Engineering
Academic title abbreviationmag. sanit. ing.
Genremaster's thesis
Language Croatian
Defense date2016-09-29
Parallel abstract (English)
Saccharomyces cerevisiae, also known as baker's or brewer's yeast has been used for centuries in the beer, wine and bread industry. Today, it is one of the most widely used eukaryotic model organism, especially to study the effects of environmental toxicants such as iron. Iron is essential element for biological systems but in excessive amounts can cause ROS production and consequently cell damage. The aim of this work was to analyze the effect of high iron concentration on proteome and lipidome of the yeast Saccharomyces cerevisiae W303 mitochondria. Research was carried out on a yeast grown in a media suplemented with iron and on yeast grown in unsuplemented media used as a control sample. Proteins of isolated yeast mitochondria were separated using 2-D electrophoresis, stained with Sypro Ruby solution and visualized using the UV transillumination. Protein profiles of treated and untreated cells were compared using Dymension software from Syngene. The number of detected proteins was lower in sample treated with iron. 9 proteins were common to both samples, eight of which were up-regulated and one was down-regulated. Also, the exposure to Fe3+ ions resulted in de novo synthesis of 5 proteins. Two proteins were detected using Protein Pilot v4.5 program: subunit of KGDH, enzyme involved in the Krebs cycle and CPR6, member of the cyclophilin family. The changes in lipidome level were determined by analysis of fatty acid composition using gas chromatography. The major changes in mitochondrial lipids of treated cells were decrease in fatty acid unsaturation and increase in chain length. Lipidomic analysis also revealed reduction of the lipid to protein mass ratio as a response to eleveted iron.
Parallel keywords (Croatian)kvasac željezo mitohondriji
Resource typetext
Access conditionAccess restricted to students and staff of home institution
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:184:921489
CommitterBosa Licul