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master's thesis
Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori
Zagreb: University of Zagreb, Faculty of Science, 2019. urn:nbn:hr:217:415997
Faletar, Mihovil
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

Cite this document

Faletar, M. (2019). Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:415997

Faletar, Mihovil. "Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:415997

Faletar, Mihovil. "Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori." Master's thesis, University of Zagreb, Faculty of Science, 2019. https://urn.nsk.hr/urn:nbn:hr:217:415997

Faletar, M. (2019). 'Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori', Master's thesis, University of Zagreb, Faculty of Science, accessed 20 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:415997

Faletar M. Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2019 [cited 2024 November 20] Available at: https://urn.nsk.hr/urn:nbn:hr:217:415997

M. Faletar, "Određivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2019. Available at: https://urn.nsk.hr/urn:nbn:hr:217:415997

Metadata
TitleOdređivanje interakcije bakrovih iona i proteina CrdA iz bakterije Helicobacter pylori
Title (english)Determination of the interaction of copper ions and CrdA protein from Helicobacter pylori
AuthorMihovil Faletar
MentorDubravka Matković-Čalogović (mentor)
Committee memberDubravka Matković-Čalogović (predsjednik povjerenstva)
Committee memberMorana Dulić (član povjerenstva)
Committee memberIva Juranović Cindrić (član povjerenstva)
Committee memberBiserka Prugovečki (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2019-09-30, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
Proučavan je protein CrdA humanog patogena bakterije Helicobacter pylori. Smatra se da protein CrdA sudjeluje u transportu iona bakra. U svrhu ispitivanja mehanizma regulacije iona bakra u H. pylori i dobivanja proteina CrdA provedena je prekomjerna ekspresija dva fuzijska proteina SUMO-G-CrdA i GST-CrdA u bakteriji Escherichia coli. Provedena su pročišćavanja fuzijskih i porezanih proteina pomoću afinitetne kromatografije, gel-filtracije i ionskoizmjenjivačke kromatografije. Optimizirane su metode rezanja i pročišćavanja u svrhu dobivanja što čišćih uzoraka proteina CrdA. Ispitana je stabilnost fuzijskog proteina SUMO-GCrdA mjerenjem temperature denaturacije proteina pri različitim uvjetima kao i DLS metodom. Ustanovljeno je da je najveća stabilnost fuzijskog proteina u otopini koja sadrži Cu2+ ione što bi moglo upućivati na stvaranje interakcije između proteina CrdA i Cu2+ iona. Ispitivanjem interakcije proteina CrdA i Cu2+ iona mikrotermoforezom određena je konstanta disocijacije kompleksa CrdA:Cu2+ koja iznosi 7,98 (± 1,94) μmol dm-3.
Abstract (english)
Protein CrdA from the human pathogen Helicobacter pylori was studied. The CrdA protein is thought to participate in the transport of copper ions. In order to investigate the mechanism of copper ion regulation in H. pylori and to obtain the CrdA protein, overexpression of two fusion proteins SUMO-G-CrdA and GST-CrdA was performed in the Escherichia coli. Purification of fusion and cleaved proteins was performed using affinity chromatography, gel-filtration and ion-exchange chromatography. Cleavage and purification were optimized to obtain pure CrdA protein samples. Stability of the fusion protein SUMO-G-CrdA was tested by measuring change in the thermal denaturation temperature under varying conditions and by the DLS method. The highest stability of the fusion protein was found to be in the Cu2+ solution suggesting a possible interaction between the CrdA protein and Cu2+ ions. The protein complex CrdA:Cu2+ dissociation constant was determined by microthermophoresis and its value was determined to be 7.98 (± 1.94) μmol dm−3.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:415997
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 on2019-10-22 11:40:04