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master's thesis
Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse
Zagreb: University of Zagreb, Faculty of Science, 2023. urn:nbn:hr:217:251991
Kindl, Domagoj Christian
University of Zagreb
Faculty of Science
Department of Biology
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

Cite this document

Kindl, D. C. (2023). Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse (Master's thesis). Zagreb: University of Zagreb, Faculty of Science; Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:251991

Kindl, Domagoj Christian. "Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse." Master's thesis, University of Zagreb, Faculty of Science; University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:251991

Kindl, Domagoj Christian. "Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse." Master's thesis, University of Zagreb, Faculty of Science; University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:251991

Kindl, D. C. (2023). 'Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse', Master's thesis, University of Zagreb, Faculty of Science; University of Zagreb, Faculty of Science, accessed 13 May 2024, https://urn.nsk.hr/urn:nbn:hr:217:251991

Kindl DC. Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; Zagreb: University of Zagreb, Faculty of Science; 2023 [cited 2024 May 13] Available at: https://urn.nsk.hr/urn:nbn:hr:217:251991

D. C. Kindl, "Istraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse", Master's thesis, University of Zagreb, Faculty of Science, Zagreb; University of Zagreb, Faculty of Science, Zagreb, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:217:251991

Metadata
TitleIstraživanje biokemijskih osobina mutiranih inačica dipeptidil - peptidaze III, R510W i V514M te njihovog utjecaja na regulaciju signalnog puta Nrf2/Keap1 ; Enzimi - primjer dobre nastavne prakse
Title (english)Investigation of the biochemical properties of R510W and V514M mutant variants of dipeptidyl peptidase III and their impact on the regulation of Nrf2/Keap1 signalling pathway ; Enzymes - an example of good teaching practice
AuthorDomagoj Christian Kindl
MentorMihaela Matovina (mentor)
MentorDraginja Mrvoš-Sermek (mentor)
Committee memberDraginja Mrvoš-Sermek (predsjednik povjerenstva)
Committee memberJasmina Rokov Plavec (član povjerenstva)
Committee memberInes Radanović (član povjerenstva)
Committee memberNada Oršolić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2023-09-28, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
Diplomski rad sastoji se od eksperimentalnog i metodičkog dijela. U eksperimentalnom dijelu ispitane su biokemijske osobine mutiranih inačica R510W i V514M humane dipeptidil– peptidaze III (hDPP III) čije su genomske sekvence nađene u uzorcima zloćudnih tumora, te je ispitan njihov utjecaj na regulaciju signalnog puta Nrf2/Keap1. Ciljanom mutagenezom napravljeni su vektori za ekspresiju mutiranih inačica proteina te su one eksprimirane u bakteriji Escherichia coli i pročišćene. Utvrđeno je da je aktivnost svih triju proteina viša u prisutnosti kobaltovih iona, a analizom kinetike reakcije je utvrđeno da R510W ima oko 1000 puta, a V514M oko 2 puta nižu katalitičku aktivnost od divljeg tipa. Određivanjem razine ekspresije gena utvrđeno je da prekomjerna ekspresija divljeg tipa i mutiranih inačica hDPP III nemaju utjecaj na ekspresiju gena kontroliranih s Nrf2 u HEK293T stanicama. U sklopu metodičkog dijela rada osmišljeni su pokusi na temu enzima, temeljem kojih su oblikovani metodičko-didaktički materijali za jedan 90-minutni nastavni sat. Pri tome su postavljeni odgojno-obrazovni ciljevi temeljeni na prethodnim učeničkim znanjima, vještinama i sposobnostima.
Abstract (english)
The thesis consists of an experimental and methodological part. In the experimental part, the biochemical properties of the mutant variants of human dipeptidyl peptidase III (hDPP III), whose genomic sequences were found in malignant tumors, R510W i V514M, and their influence on the regulation of the Nrf2/Keap1 signaling pathway were examined. The expression vectors for the mutants were constructed using site-directed mutagenesis and expressed in E. coli and purified. The activity of all three proteins was higher in the presence of cobalt ions, and the kinetic parameters analysis revealed that R510W has about 1000 times and V514M about 1 times lower catalytic activity than the wild type. Gene expression analysis detected that overexpression of the wild-type and mutants of hDPP III had no effect on the expression of genes controlled by Nrf2 in HEK293T cells. In the methodological part of the thesis, experiments on the topic of enzymes as well as methodical-didactic materials related to 90-minute lesson were designed. At the same time, educational goals were set based on the students' previous knowledge, skills and competencies.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:251991
ProjectNumber: IP-2018-01-2936
Title: Biološka važnost dipeptidil peptidaze III i njezin utjecaj na zdravlje čovjeka
Acronym: DPP3BioRe
Leader: Sanja Tomić
Jurisdiction: Croatia
Funder: HRZZ
Funding stream: IP
Study programmeTitle: Integrated undergraduate and graduate university study of Biology and Chemistry Education; specializations in: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Course: Integrated undergraduate and graduate university study of Biology and Chemistry Education
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra edukacije biologije i kemije (magistar/magistra edukacije biologije i kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2023-10-27 12:31:07