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master's thesis
Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu
Zagreb: University of Zagreb, Faculty of Science, 2019. urn:nbn:hr:217:995612
Brkić, Alojzije
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

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Brkić, A. (2019). Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:995612

Brkić, Alojzije. "Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:995612

Brkić, Alojzije. "Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:995612

Brkić, A. (2019). 'Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:995612

Brkić A. Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2019 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:995612

A. Brkić, "Utjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:217:995612

Metadata
TitleUtjecaj mistranslacije na stabilnost modelnih proteina u staničnom ekstraktu
Title (english)Effect of mistranslation on the stability of model proteins in cell extract
AuthorAlojzije Brkić
MentorIta Gruić Sovulj (mentor)
MentorMarko Močibob (mentor)
Committee memberMarko Močibob (predsjednik povjerenstva)
Committee memberIvica Đilović (član povjerenstva)
Committee memberSnežana Miljanić (član povjerenstva)
Committee memberIva Juranović Cindrić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2019-02-25, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
Izoleucil-tRNA-sintetaza (IleRS) katalizira sintezu Ile-tRNAIle u procesu biosinteze proteina. IleRS može ponekad griješiti i misacilirati tRNAIle s valinom ili neproteinogenim norvalinom. Pogreška u biosintezi proteina (mistranslacija) može biti štetna za stanicu. Iz tog razloga, IleRS posjeduje domenu za popravak vlastite pogreške. U okviru ovog rada praćena je promjena temperature mekšanja β-galaktozidaze kao modelnog proteina uslijed zamjene izoleucina s valinom ili norvalinom. Termička stabilnost β-galaktozidaze praćena je u staničnim ekstraktima soja bakterije Escherichia coli koji ima inaktiviranu domenu za popravak pogreške IleRS te ugrađuje valin i norvalin na izoleucinske položaje. Pokazano je da mistranslacija izoleucinskih položaja β-galaktozidaze objema nepripadnim aminokiselinama smanjuje termičku stabilnost enzima te da je utjecaj norvalina izraženiji. Nadalje, opažena je značajnija inhibicija rasta bakterijskih stanica uslijed zamjene izoleucina norvalinom u odnosu na valin. Rezultati sugeriraju da veća toksičnost mistranslacije norvalinom barem djelomično potječe od smanjene stabilnosti proteina s ugrađenim norvalinom.
Abstract (english)
Isoleucyl-tRNA synthetase (IleRS) catalyses formation of Ile-tRNAIle for protein biosynthesis. IleRS may mistakingly activate tRNAIle with valine and nonproteinogenic norvaline. However, IleRS uses an editing domain and corrects its own mistakes to prevent accumulation of erroneous proteins. This work focused on the effects of isoleucine to valine or isoleucine to norvaline substitutions on the thermal stability of a model protein, β-galactosidase. The melting point of β-galactosidase was measured in cell extracts of an IleRS editing-deficient strain of Escherichia coli, which incorporated valine or norvaline at isoleucine positions. We showed that mistranslation of β-galactosidase with both norvaline and valine lowered the thermal stability of the enzyme, with norvaline introducing a higher effect. Besides that, isoleucine to norvaline substitution inhibited the growth of the editing-deficient strain to a higher extent than isoleucine replacement with valine. The results suggest that higher toxicity of norvaline misincorporation may be in part caused by production of norvalylated proteins with a lower thermal stability.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:995612
Study programmeTitle: Graduate university studies in chemistry; specializations in: Research programme
Course: Research programme
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2019-02-28 11:25:41