Mapiranje malih GTPaza HRAS, NRAS i MRAS korištenjem metode ligacije proteina posredovane pocijepanim inteinom

master's thesis

• Title (english): Mapping of small GTPase proteins HRAS, NRAS and MRAS using the Split Intein-Mediated Protein Ligation assay
• Author: Josipa Mateševac
• Mentor: Petra Korać (mentor)
• Committee member: Jasna Lajtner (predsjednik povjerenstva)
• Committee member: Sofia Ana Blažević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2020-07-17, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Nadporodica malih GTPaza Ras ima ulogu u prijenosu signala i modulaciji staničnih procesa.
Njeni članovi djeluju kao binarni molekularni prekidači koji izmjenjuju inaktivni oblik vezan
za GDP i aktivni oblik vezan za GTP koji veže svoje nizvodne efektore s velikim afinitetom.
Porodica Ras uključena je u regulaciju unutarstanične signalizacije i utječe na procese kao što
su stanični rast, preživljavanje, morfologija i diferencijacija. Proteini RAS post-translacijski
se modificiraju adicijom lipida koja omogućava njihovo vezanje za membrane i biokemijsku
aktivnost u raznim signalnim putovima. Nepravilnosti u normalnoj funkciji proteina obično
rezultiraju malignim promjenama. Određene mutacije različitih proteina RAS često se
pronalaze u različitim vrstama tumora, a također mogu voditi i do kongenitalnih razvojnih
poremećaja. Budući da međuproteinske interakcije imaju ulogu u patofiziologiji bolesti,
njihovo je razumijevanje ključno u otkrivanju potencijalnih meta terapija. Cilj ovog
istraživanja bio je mapirati interakcije između tri proteina porodice Ras, HRAS, NRAS i
MRAS, i proteina koji su s njima potencijalno u interakciji koristeći novorazvijen pristup,
SIMPL (Split Intein-Mediated Protein Ligation), koji omogućava kvantifikaciju
međuproteinskih interakcija metodom ELISA. Osam interaktora identificirano je za HRAS,
trideset i četiri za NRAS te osamnaest za MRAS.

Abstract (english)

The Ras superfamily of small GTPases plays a role in signal transduction and modulation of
cellular processes. Its members act as binary molecular switches that cycle between the
inactive, GDP-bound form, and the active, GTP-bound form, in which they bind their
downstream effectors with high affinity. The Ras family is involved in regulation of
intracellular signaling and affects processes such as cell growth, survival, morphology and
differentiation. They are post-translationally modified by lipid addition which enables their
membrane recruitment and biochemical activity in various signaling pathways. Irregularities
in the normal function of RAS proteins usually result in malignant changes. Certain mutations
of various RAS proteins are commonly found in distinct cancer types and can also lead to
different congenital developmental disorders. As protein-protein interactions play a role in
disease pathophysiology, understanding of PPIs is crucial in discovering potential drug
therapy targets. The aim of this study was to map interactions between three members of the
Ras family, HRAS, NRAS and MRAS and their potential interactors using the newly
developed SIMPL assay which allows PPI quantification using ELISA. Eight interactors were
identified for HRAS, thirty-four for NRAS and eighteen for MRAS.

• Keywords: GTPaze; RAS; PPI; inteini; proteinski splicing; SIMPL
• Keywords (english): GTPase; RAS; PPI; inteins; protein splicing; SIMPL
• Language: croatian
• URN:NBN: urn:nbn:hr:217:135174
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2022-07-17
• Created on: 2020-07-20 11:33:57