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master's thesis
Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom
Zagreb: University of Zagreb, Faculty of Science, 2021. urn:nbn:hr:217:560473
Valla, Robert
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

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Valla, R. (2021). Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:560473

Valla, Robert. "Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:560473

Valla, Robert. "Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom." Master's thesis, University of Zagreb, Faculty of Science, 2021. https://urn.nsk.hr/urn:nbn:hr:217:560473

Valla, R. (2021). 'Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom', Master's thesis, University of Zagreb, Faculty of Science, accessed 23 December 2024, https://urn.nsk.hr/urn:nbn:hr:217:560473

Valla R. Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2021 [cited 2024 December 23] Available at: https://urn.nsk.hr/urn:nbn:hr:217:560473

R. Valla, "Karakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2021. Available at: https://urn.nsk.hr/urn:nbn:hr:217:560473

Metadata
TitleKarakterizacija interakcije proteina BLNK i receptorne tirozin-kinaze MET uporabom ligacije proteina posredovane pocijepanim inteinom
Title (english)Characterization of BLNK protein interaction with receptor tyrosine-kinase MET using split intein-mediated protein ligation
AuthorRobert Valla
MentorIgor Štagljar (mentor)
MentorNataša Bauer (sumentor)
Committee memberNataša Bauer (predsjednik povjerenstva)
Committee memberNada Oršolić (član povjerenstva)
Committee memberMarin Ježić (član povjerenstva)
Committee memberPetra Korać (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2021-07-19, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
Receptorna tirozin-kinaza MET je protoonkogen uključen u razne fiziološke procese, sudjeluje u tkivnoj homeostazi i regeneraciji te u brojnim patofiziološkim stanjima, primjerice različitim tipovima raka. Protein BLNK je središnji adaptorni protein nizvodno od signalne kaskade B-staničnog receptora. Protein BLNK omogućuje stvaranje multimolekularnih signalnih kompleksa, što prethodi sazrijevanju limfocita B. Protein BLNK je novo opisani interackijski partner receptora MET te bi interakcija receptora MET i proteina BLNK mogla imati jedinstvenu biološku značajnost. U ovom istraživanju je u svrhu karakterizacije interakcije receptora MET i proteina BLNK korištena nova metoda istraživanja proteinskih interakcija zvana ligacija proteina posredovana pocijepanim inteinom (engl. SIMPL). Interakcija receptora MET i proteina BLNK je potvrđena te je istražena ovisnost interakcije o fosforilaciji. Uspostavljena je i okarakterizirana stanična linija sa stabilnom ekpresijom receptora MET i taj se stabilni sustav MET-SIMPL može upotrijebiti kao platforma za probir lijekova. Identificirane su aminokiseline odgovorne za interakciju receptora MET i proteina BLNK. Metoda SIMPL ELISA također je upotrebljena za određivanje veznih mjesta proteina GRB2 na protein BLNK te za istraživanje uloge proteina GRB2 u interakciji receptora MET i proteina BLNK. Dobiveni rezultati pružaju bolji uvid u funkcionalne i mehanističke značajke interakcije receptora MET i proteina BLNK i proširuju razumijevanje biologije receptorne tirozin-kinaze MET.
Abstract (english)
MET receptor tyrosine-kinase is a proto-oncogene involved in various physiological processes, such as tissue homeostasis and regeneration, as well as numerous pathophysiological conditions, such as different types of cancers. BLNK is a central adaptor protein downstream of the B-cell receptor signaling cascade, which enables the formation of multimolecular signaling complexes preceding the maturation of B lymphocytes. BLNK protein is a newly discovered interaction partner of MET, and MET-BLNK interaction may have a unique biological significance. A novel method to study protein-protein interactions called split intein-mediated protein ligation (SIMPL) was used in this research to characterize MET-BLNK interaction. MET-BLNK interaction was orthogonally confirmed and its phosphorylation-dependent nature was investigated using the SIMPL assay. The stable MET-SIMPL system was generated, characterized, and utilized as a drug screening platform. The amino acid residues on MET and BLNK responsible for MET-BLNK interaction were identified using SIMPL assay. SIMPL ELISA was used to determine the binding sites for GRB2 protein on BLNK and assess the role of GRB2 in MET-BLNK interaction. These results provided insight into functional and mechanistic features of MET-BLNK interaction and have deepened the understanding of MET receptor tyrosine-kinase biology.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:217:560473
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: 2023-07-19
Terms of useLicence In copyright icon
Created on2021-07-22 12:48:06