prikaz prve stranice dokumenta Regulation of the activity of ubiquitin ligase HACE1
Embargo period 2025-09-21
master's thesis
Regulation of the activity of ubiquitin ligase HACE1
Zagreb: University of Zagreb, Faculty of Science, 2023. urn:nbn:hr:217:358497
Toplak, Julia Jelena
University of Zagreb
Faculty of Science
Department of Biology

Institutional repository: Repository of the Faculty of Science

Cite this document

Toplak, J. J. (2023). Regulation of the activity of ubiquitin ligase HACE1 (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:358497

Toplak, Julia Jelena. "Regulation of the activity of ubiquitin ligase HACE1." Master's thesis, University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:358497

Toplak, Julia Jelena. "Regulation of the activity of ubiquitin ligase HACE1." Master's thesis, University of Zagreb, Faculty of Science, 2023. https://urn.nsk.hr/urn:nbn:hr:217:358497

Toplak, J. J. (2023). 'Regulation of the activity of ubiquitin ligase HACE1', Master's thesis, University of Zagreb, Faculty of Science, accessed 25 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:358497

Toplak JJ. Regulation of the activity of ubiquitin ligase HACE1 [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2023 [cited 2024 November 25] Available at: https://urn.nsk.hr/urn:nbn:hr:217:358497

J. J. Toplak, "Regulation of the activity of ubiquitin ligase HACE1", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2023. Available at: https://urn.nsk.hr/urn:nbn:hr:217:358497

Metadata
TitleRegulation of the activity of ubiquitin ligase HACE1
Title (croatian)Regulacija aktivnosti ubikvitinske ligaze HACE1
AuthorJulia Jelena Toplak
MentorSonja Lorenz (mentor)
MentorDunja Leljak-Levanić (komentor)
Committee memberDunja Leljak-Levanić (predsjednik povjerenstva)
Committee memberBiljana Balen (član povjerenstva)
Committee memberIvana Ivančić Baće (član povjerenstva)
Committee memberPetra Peharec Štefanić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Biology)
Zagreb
Defense date and country2023-09-21, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
The HECT ubiquitin ligase HACE1 is involved in membrane dynamics regulation, mitophagy, redox homeostasis, and cytoskeleton organization. Despite HACE1 being a tumor suppressor and a significant factor in neural development, it remains biochemically poorly characterized, and its regulatory mechanisms are not known. This master’s thesis explores the impact of phosphorylation as a regulatory mechanism and its effect on HACE1 activity and dimerization. Phosphomimetic variants were generated and characterized using functional assays, mass photometry, and analytical size exclusion chromatography. The findings reveal that the phosphomimetic mutations within the N-terminal helix disrupted dimerization and activated HACE1 activity, providing insights into how the monomer could be stabilized in its active state. Furthermore, recombinantly purified kinases PAK1 and TBK1 were investigated as potential regulators of HACE1 activity, and the sites targeted by these kinases in HACE1 sites were identified through mass spectrometry.
Abstract (croatian)
Ubikvitinska HECT ligaza HACE1 sudjeluje u regulacije dinamike membrana, mitofagiji, redokshomeostazi i organizaciji citoskeleta. Unatoč tome što je ligaza HACE1 tumor-supresor i značajan faktor u razvoju živčanog sustava, nije detaljno biokemijski karakterizirana, a regulatorni mehanizmi nisu poznati. Ovaj diplomski rad istražuje utjecaj fosforilacije kao regulatornog mehanizma na aktivnost i dimerizaciju HACE1. Pripremljene su fosfomimetičke varijante i karakterizirane korištenjem funkcionalnih eseja, masene fotometrije i analitičke kromatografije temeljene na isključivanju. Rezultati otkrivaju da fosfomimetičke varijante unutar N-terminalnog heliksa narušavaju dimerizaciju i aktiviraju aktivnost ligaze HACE1, pružajući uvid u to kako se monomer može stabilizirati u aktivnom stanju. Nadalje, rekombinantno pročišćene kinaze PAK1 i TBK1 istražene su kao potencijalni regulatori. Mjesta u ligazi HACE1 koje te kinaze ciljaju identificirana su putem masene spektrometrije.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:217:358497
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 conditionsEmbargoed access
Embargo expiration date: 2025-09-21
Terms of useLicence In copyright icon
Created on2023-10-25 10:22:08