Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori

Petek, Ana

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master's thesis

Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori

Zagreb: University of Zagreb, Faculty of Science, 2020. urn:nbn:hr:217:562036

Petek, Ana

University of Zagreb
Faculty of Science
Department of Chemistry

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APA 6th Edition

Petek, A. (2020). Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:562036

MLA 8th Edition

Petek, Ana. "Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori." Master's thesis, University of Zagreb, Faculty of Science, 2020. https://urn.nsk.hr/urn:nbn:hr:217:562036

Chicago 17th Edition

Harvard

Petek, A. (2020). 'Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori', Master's thesis, University of Zagreb, Faculty of Science, accessed 27 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:562036

Vancouver

Petek A. Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2020 [cited 2024 December 27] Available at: https://urn.nsk.hr/urn:nbn:hr:217:562036

IEEE

A. Petek, "Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2020. Available at: https://urn.nsk.hr/urn:nbn:hr:217:562036

Cite this item: https://urn.nsk.hr/urn:nbn:hr:217:562036

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Title	Utjecaj afinitetnog privjeska na svojstva adenilosukcinat-sintetaze iz bakterije Helicobacter pylori
Title (english)	Effect of affinity tag on properties of adenylosuccinate synthetase from Helicobacter pylori
Author	Ana Petek
Mentor	Ivana Leščić Ašler (mentor)
Committee member	Branimir Bertoša (predsjednik povjerenstva)
Committee member	Jasmina Rokov Plavec (član povjerenstva)
Committee member	Iva Juranović Cindrić (član povjerenstva)
Committee member	Nikola Bregović (član povjerenstva)
Granter	University of Zagreb Faculty of Science (Department of Chemistry) Zagreb
Defense date and country	2020-09-29, Croatia
Scientific / art field, discipline and subdiscipline	NATURAL SCIENCES Chemistry
Abstract	Bakterija Helicobacter pylori za sintezu purina koristi reciklirajući put, a ključni enzim u tom putu jest adenilosukcinat-sintetaza (AdSS). Histidinski afinitetni privjesak omogućuje jednostavnije pročišćavanje proteina uz veći prinos, no istovremeno može utjecati na prvobitnu funkciju enzima. Stoga je provedena usporedba nativnog AdSS-a i AdSS-a obilježenog histidinskim privjeskom, kao cilj ovog diplomskog rada. Obilježeni AdSS je pročišćen u dva koraka, afinitetnom kromatografijom i gel filtracijom. Izmjerena mu je specifična enzimska aktivnost, te su određeni kinetički parametri za sva tri supstrata reakcije. Provedena je inhibicija AdSS-a jednim od produkata reakcije – GDP-om. Određeno je temperaturno i pH područje stabilnosti ovog enzima. Udio elemenata sekundarne strukture u AdSS-u određen je metodom cirkularnog dikroizma. Enzim je kristaliziran tehnikom difuzije para iz viseće kapi, pri različitim koncentracijama proteina. Dobiveni rezultati pokazuju kako histidinski privjesak ne utječe znatno na funkciju AdSS, te se tako obilježeni enzim može pouzdano koristiti u budućim istraživanjima.
Abstract (english)	The bacterium Helicobacter pylori utilises the recycling pathway for purine synthesis, and the key enzyme of this pathway is adenylosuccinate synthetase (AdSS). The histidine affinity tag makes protein purification easier and protein yield higher but simultaneously it can affect the original function of the enzyme. Therefore, a comparison of native and histidine tagged AdSS was conducted as the aim of this thesis. The tagged AdSS was purified in two steps, by affinity chromatography and gel filtration. Its specific enzyme activity was measured and the kinetic parameters for all three reaction substrates were determined. Inhibition of AdSS was performed by one of the reaction products – GDP. The temperature and pH range of stability of this enzyme was determined. The content of the secondary structure elements in AdSS was determined by the circular dichroism method. The enzyme was crystallized by hanging drop vapor diffusion technique at different protein concentrations. Obtained results showed that histidine tag does not affect function of AdSS significantly so tagged enzyme can be used reliably in the future research.
Keywords
Keywords (english)
Language	croatian
URN:NBN	urn:nbn:hr:217:562036
Study programme	Title: 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 resource	Text
File origin	Born digital
Access conditions	Open access
Terms of use
Repository	Repository of the Faculty of Science
Created on	2020-10-09 09:22:56