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master's thesis
Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora
Zagreb: University of Zagreb, Faculty of Science, 2018. urn:nbn:hr:217:896663
Pipalović, Goran
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

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Pipalović, G. (2018). Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:896663

Pipalović, Goran. "Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:896663

Pipalović, Goran. "Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:896663

Pipalović, G. (2018). 'Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora', Master's thesis, University of Zagreb, Faculty of Science, accessed 21 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:896663

Pipalović G. Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2018 [cited 2024 December 21] Available at: https://urn.nsk.hr/urn:nbn:hr:217:896663

G. Pipalović, "Prekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2018. Available at: https://urn.nsk.hr/urn:nbn:hr:217:896663

Metadata
TitlePrekomjerna ekspresija i biokemijska karakterizacija mutanata humane dipeptidil-peptidaze III iz zloćudnih tumora
Title (english)Overexpression and biochemical characterization of human dipeptidyl-peptidase III mutants found in cancer
AuthorGoran Pipalović
MentorMihaela Matovina (mentor)
Committee memberAleksandra Maršavelski (predsjednik povjerenstva)
Committee memberŽeljka Soldin (član povjerenstva)
Committee memberVesna Petrović Peroković (član povjerenstva)
Committee memberMorana Dulić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2018-09-27, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
Dipeptidil-peptidaze III (DPP III) su metalopeptidaze iz porodice M49 koje hidroliziraju dipeptide s N-kraja peptidnih supstrata veličine 3 do 10 aminokiselina. Fiziološka funkcija ove peptidaze nije još do kraja razjašnjena, ali pretpostavlja se da ljudska DPP III osim uloge u završnim koracima razgradnje peptida u stanici ima ulogu i u nekoliko patofizioloških procesa, uključujući razvitak raka. U cBioPortal bazi podataka nalaze se sekvencirani uzorci DNA izolirane iz ljudskih tumora te su nađene mutacije u kodirajućoj regiji gena za humanu DPP III. Odabrane su tri mutacije kako bi se ispitao učinak istih na biokemijska svojstva i aktivnost humane DPP III. Istraživanje se provodilo na mutantima humane DPP III: E451K, R638L i R638W koji su dobiveni ciljanom in vitro mutagenezom ekspresijskih plazmida. Nakon tog slijedila je ekspresija mutanata u bakteriji E. coli te njihova izolacija i pročišćavanje afinitetnom kromatografijom na Ni-NTA koloni. Mutanti su podvrgnuti raznim testovima aktivnosti i mjerenju kinetike te su analizirani nativnom elektroforezom i izoelektričnim fokusiranjem na gelu. Dobiveni parametri za mutante uspoređeni su s onima „divljeg“ tipa.
Abstract (english)
Dipeptidyl peptidases III (DPP III) are members of the M49 metalopeptidase family which hydrolyze dipeptides from the N-termini of 3 to 10 aminoacids long peptides. Their physiological function is not yet fully understood, but it is assumed that the human DPP III, apart from the role in the final steps of peptide degradation in the cell, has the role in several pathophysiological processes, including the development of cancer. The cBioPortal database contains sequenced DNA samples isolated from human cancers and mutations in the coding region of the human DPP III gene are found. Three mutations were selected to examine their effect on the biochemical properties and the activity of human DPP III. The study was performed on human DPP III: E451K, R638L and R638W mutants obtained by site-directed in vitro mutagenesis of expression plasmids, followed by the expression of mutants in E. coli, and the isolation and purification by affinity chromatography on Ni-NTA column. The mutants were subjected to various activity assays, kinetics measurement, and they were also analysed by native and isoelectric focusing gel electrophoresis. The obtained mutant parameters were compared to those of the wild type.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:896663
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
Embargo expiration date: 2020-09-27
Terms of useLicence In copyright icon
Created on2018-11-23 11:47:47