master's thesis
Utjecaj iona olova Pb2+ na mitohondrije kvasca Saccharomyces cerevisiae W303

Nermina Mumiši (2016)
Sveučilište u Rijeci
Medicinski fakultet
Katedra za kemiju i biokemiju
Metadata
TitleUtjecaj iona olova Pb2+ na mitohondrije kvasca Saccharomyces cerevisiae W303
AuthorNermina Mumiši
Mentor(s)Gordana Čanadi Jurešić (thesis advisor)
Abstract
Industrijalizacija je dovela do povećane upotrebe konzerviranih namirnica, boja, baterija i akumulatora u kojima se nalaze velike koncentracije olova. Čak i male koncentracije su štetne jer se olovo putem zraka i hrane apsorbira i kumulira u organima (kosti, jetra) i to najviše u središnjem živčanom sustavu. Olovo, kao teški metal, stvara veliku količinu ROS-a te posljedično uzrokuju promjene na proteinima i lipidima.. U ovom radu istražili smo utjecaj olovovih (II) iona, Pb2+, na mitohondrije kvasca. Pri tom se kao modelni organizam koristio kvasac Saccharomyces cerevisiae, poznat kao pivski kvasac ili pekarski kvasac, zbog jednostavnosti uzgoja i sekvencioniranog genoma. U istraživanju je korišten kvasac uzgojen na YPD podlozi s dodatkom olovovih iona u koncentraciji od 8 mmol/L i inkubiran paralelno s netretiranim kvascem koji je služio kao kontrola. Inhibicija na rast kvasca iznosila je 93,8%. Glavne masne kiseline u svim uzorcima bile su oleinska i palmitoleinska, a zatim miristinska, palmitinska i stearinska kiselina. Analiza lipida pokazala je ekspresiju masenog udjela proteina prema proteinima u mitohondrijima kvasca tretiranog olovom. Proteini mitohondrija izolirani su dvodimenzionalnom elektroforezom i obojeni dvjema metodama (bojanje s Sypro Ruby i sa srebrom). Proteinski profili tretiranih uzoraka sa Pb2+ i netretiranih uzoraka uspoređeni su kroz njihov volumen. Proteini su analizirani MS MALDI – TOF/TOF tehnikom i identificirani pomoću nrNCBI baze podataka. Od zajedničkih proteina, 25 je pokazalo značajni izričaj (p<0,05). Njih 13 pokazalo je povećani (> 2), a 12 smanjeni izričaj proteina (< -2). Identificirano je 7 proteina uključenih u ciklus limunske kiseline, sintezu proteina i transport molekula.
Keywordsyeast mitochondria lead
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 date2016-09-30
Parallel abstract (English)
Industrialization increased the amount of canned foods, paints, electrical devices, and the need to use batteries, accumulator and other products that contain high levels of lead. Even low concentrations are toxic to humans, because the lead is absorbed through the air and food and accumulated in organs (bones, liver), mostly in the central nervous system. Lead, a heavy metal, produce a large amount of ROS and consequently cause changes in proteins and lipids. The aim of this research was to study the effect of lead-ions on the yeast mitochondria. Saccharomyces cerevisiae yeast, also known as baker's or brewer's yeast was used, because it is simple to grow and manipulate and has sequenced genome. Yeast was cultivated in a YPD media with 8 mmol/L of lead ion. Untreated yeast was used as a control sample. Inhibition of the cell growth was 93,8%. The main fatty acids in all samples were oleic and palmitoleic, followed by myristic, palmitic and stearic acids. Lipidomic analysis revealed expression of the lipid to protein mass ratio induced by lead ions. Proteins from mitchondria were separated using 2D electrophoresis. Proteins from mitochondria were stained with 2 methods (staining with Sypro Ruby solution and with silver) and processed. Proteins profiles of treated (with Pb2+) and untreated cells were compared by the ratio of volume in proteins. The proteins were analyzed by MS MALDI - TOF/TOF technique and identified by nrNCBI database. 25 of matched proteins were significantly changed (p<0,05). 13 of matched proteins were significantly up- regulated (> 2 fold) and 12 were down- regulated (< -2 fold). 7 proteins involved in the citric acid cycle, protein synthesis and transport of molecules, were identified.
Parallel keywords (Croatian)kvasac mitohondrij olovo
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:184:807309
CommitterBosa Licul