prikaz prve stranice dokumenta Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum
  Download
PDF 863.19 KB
master's thesis
Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum
Zagreb: University of Zagreb, Faculty of Science, 2022. urn:nbn:hr:217:122699
Ćutić, Tamara
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Ćutić, T. (2022). Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:122699

Ćutić, Tamara. "Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:122699

Ćutić, Tamara. "Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum." Master's thesis, University of Zagreb, Faculty of Science, 2022. https://urn.nsk.hr/urn:nbn:hr:217:122699

Ćutić, T. (2022). 'Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum', Master's thesis, University of Zagreb, Faculty of Science, accessed 04 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:122699

Ćutić T. Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2022 [cited 2024 November 04] Available at: https://urn.nsk.hr/urn:nbn:hr:217:122699

T. Ćutić, "Interakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:217:122699

Metadata
TitleInterakcija proteina IqgC i Rab5A u amebi Dictyostelium discoideum
Title (english)Interaction between Dictyostelium discoideum proteins IqgC and Rab5A
AuthorTamara Ćutić
MentorVedrana Filić Mileta (mentor)
MentorIvana Ivančić Baće (sumentor)
Committee memberIvana Ivančić Baće (predsjednik povjerenstva)
Committee memberBiljana Balen (član povjerenstva)
Committee memberPetra Peharec Štefanić (član povjerenstva)
Committee memberMaja Matulić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2022-02-22, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Protein IqgC iz amebe Dictyostelium discoideum sudjeluje u makropinocitozi, neselektivnom unosu izvanstanične tekućine, kao negativni regulator proteina RasG. Protein RasG je mala GTPaza koju protein IqgC inaktivira poticanjem hidrolize vezanog GTP-a, odnosno IqgC djeluje kao njen GAP (engl. GTPase activating protein). Proteini IqgC i RasG su tijekom makropinocitoze smješteni na makropinocitotsku čašicu i novonastali makropinosom, ali IqgC ostaje na makropinosomu i nakon što je RasG disocirao. To sugerira da IqgC na makropinosomu zadržava interakcija s drugim proteinskim i/ili fosfolipidnim partnerom. Iz objavljenog interaktoma IqgC probrani su potencijalni proteini kandidati na temelju njihove uloge u sazrijevanju ranog makropinosoma te su preliminarni pokusi pokazali da se mala GTPaza Rab5A nalazi u kompleksu s IqgC. Cilj ovog rada bio je istražiti je li interakcija proteina IqgC i Rab5A izravna te usporediti prostorno-vremenske raspodjele ova dva proteina tijekom makropinocitoze. Metodom GST-pull-down potvrdili smo da se pročišćeni protein IqgC izravno veže na Rab5A. Koristeći konfokalnu mikroskopiju, pokazali smo da IqgC i Rab5A kolokaliziraju na makropinosomima, pri čemu IqgC disocira s vezikule znatno prije nego što Rab5A dosegne maksimalni intenzitet. Za razjašnjavanje fiziološke uloge ove interakcije potrebna su daljnja istraživanja.
Abstract (english)
Dictyostelium discoideum IqgC protein interacts with the small GTPase RasG and promotes its GTP-hydrolytic activity, i.e. acts as its GAP (GTPase activating protein). By suppressing Ras activity, it negatively regulates macropinocytosis, the bulk uptake of extracellular fluid. During macropinocytosis, both IqgC and RasG localize to macropinocytic cup and newly formed macropinosome, but IqgC remains on the nascent macropinosome even after RasG dissociated. This suggests that another protein and/or phospholipid interactor engages with IqgC on the macropinosome. Few potential protein candidates have been selected from the published IqgC interactome, based on their known functions during early endosome maturation. Among them was the small GTPase Rab5A that was shown to exist in a complex with IqgC. In this work, we aimed to explore whether this interaction between IqgC and Rab5A is direct and to compare spatio-temporal distributions of these proteins during macropinocytosis. We performed a GSTRab5A-pull-down assay and confirmed that purified IqgC binds directly to Rab5A. Using confocal microscopy, we demonstrated colocalization of Rab5A and IqgC on macropinosomes with IqgC leaving the vesicle well before Rab5A reached maximal intensity. Deciphering the physiological significance of this interaction requires further investigation.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:122699
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: 2023-08-22
Terms of useLicence In copyright icon
Created on2022-03-23 13:16:39