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master's thesis
Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum
Zagreb: University of Zagreb, Faculty of Science, 2018. urn:nbn:hr:217:585696
Mijanović, Lucija
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Mijanović, L. (2018). Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:585696

Mijanović, Lucija. "Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:585696

Mijanović, Lucija. "Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum." Master's thesis, University of Zagreb, Faculty of Science, 2018. https://urn.nsk.hr/urn:nbn:hr:217:585696

Mijanović, L. (2018). 'Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum', Master's thesis, University of Zagreb, Faculty of Science, accessed 04 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:585696

Mijanović L. Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2018 [cited 2024 November 04] Available at: https://urn.nsk.hr/urn:nbn:hr:217:585696

L. Mijanović, "Biokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2018. Available at: https://urn.nsk.hr/urn:nbn:hr:217:585696

Metadata
TitleBiokemijska karakterizacija interakcije proteina IqgC i male GTPaze RASG iz amebe Dictyostelium discoideum
Title (english)Biochemical characterization of the interaction between Dictyostelium IQGAP-related protein IqgC and small GTPase RasG
AuthorLucija Mijanović
MentorVedrana Filić Mileta (mentor)
MentorIvana Ivančić Baće (mentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberJasna Hrenović (član povjerenstva)
Committee memberDamjan Franjević (član povjerenstva)
Committee memberBiljana Balen (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2018-02-23, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Proteini IQGAP u pravilu ne vežu proteine Ras i ne posjeduju sposobnost poticanja GTPazne akivnosti malih GTPaza Ras (aktivnost RasGAP). Međutim preliminarni rezultati karakterizacije Dictyostelium proteina IqgC, koji je srodan proteinima IQGAP sisavaca, sugerirali su aktivnost RasGAP ovog proteina prema Dictyostelium GTPazi RasG. Cilj ovog rada bio je biokemijskim metodama demonstrirati interakciju proteina IqgC i RasG te ispitati posjeduje li IqgC aktivnost GAP prema GTPazi RasG. Interakcija je potvrđena metodama pull-down, ko-imunoprecipitacija i bimolekularna komplementacija fluorescencije, nakon čega je napravljen test aktivacije proteina RasG koji je sugerirao aktivnost GAP. Konačna potvrda da IqgC ima aktivnost RasGAP dobivena je testom GAP s pročišćenim proteinima in vitro. U testu su korišteni ljudska GTPaza H-Ras i Dictyostelium GTPaza RasG te je pokazano da IqgC potiče, inače slabu intrinzičnu, hidrolizu GTP-a obaju GTPaza. Ovi eksperimenti nedvosmisleno pokazuju da je IqgC netipičan srodnik proteina IQGAP, koji za razliku od druga dva predstavnika u organizmu D. discoideum, DGAP1 i GAPA, kao i ostalih pripadnika ove obitelji u viših eukariota, jer osim što veže Ras stimulira i njegovu GTPaznu aktivnost time ga deaktivirajući u stanici.
Abstract (english)
IQGAP proteins generally do not interact with small GTPases from the Ras family nor stimulate their GTPase activity as do RasGAPs (Ras GTPase activating proteins). However, preliminary characterization of Dictyostelium IQGAP-related protein IqgC suggested it could have GAP activity toward the Dictyostelium GTPase RasG. The aim of this study was to biochemically validate the interaction between IqgC and RasG and to test whether IqgC exerts GAP activity toward the RasG GTPase. Pull down, co-immunoprecipitation and bimolecular fluorescence complementation were methods used to confirm the interaction, followed by Ras activation assay which indicated that IqgC has GAP activity. Finally, in vitro GAP assay, in which purified human GTPase H-Ras and Dictyostelium GTPase RasG were tested, showed that IqgC accelerates otherwise weak intrinsic GTP hidrolysis of both H-Ras and RasG. These experiments unequivocally demonstrate that this atypical IQGAP-related protein is a genuine RasGAP. In contrast to the other two Dyctiostelium IQGAPs, DGAP1 and GAPA, as well as other IQGAPs, IqgC not only binds Ras but also stimulates its GTPase activity thus deactivating Ras in the cell.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:585696
Study programmeTitle: Molecular Biology
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Embargo expiration date: 2019-02-23
Terms of useLicence In copyright icon
Created on2018-03-16 09:24:13