20.500.12556/RUL-187927 Identifikacija vezavnih partnerjev rastlinskega analoga hormona T3 med proteini navadnega repnjakovca Identification of binding partners for the plant analog of the T3 hormone among Arabidopsis thaliana proteins Ščitnična hormona trijodotironin (T3) in tiroksin oziroma tetrajodotironin (T4) imata pri človeku in drugih živalih pomembno vlogo pri uravnavanju razvoja, rasti in metabolizma. V zadnjih letih se pojavlja vse več dokazov, da so spojine, podobne ščitničnim hormonom, prisotne tudi v rastlinah, vendar njihova fiziološka vloga in molekularni mehanizmi delovanja še niso povsem pojasnjeni. Namen magistrske naloge je bil v modelni rastlini navadni repnjakovec (Arabidopsis thaliana) identificirati potencialne proteine, ki vežejo hormon T3. V okviru eksperimentalnega dela smo T3 imobilizirali na magnetne kroglice in izvedli test »pull down« z uporabo lizata rastline A. thaliana. Elucijske frakcije smo poslali na analizo z masno spektrometrijo, identificirane proteine pa funkcionalno razvrstili na podlagi podatkov iz podatkovne zbirke UniProt. Poleg tega smo pripravili fluorescenčno označen T3 z barvilom Alexa Fluor 488, ga inkubirali z rastlinskim lizatom ter z nativno gelsko elektroforezo analizirali vezavo označenega hormona na rastlinske proteine. Analiza z masno spektrometrijo je omogočila identifikacijo več kot sto proteinov, med katerimi smo z uporabo različnih statističnih pristopov določili osem najbolj prepričljivih kandidatov. Med njimi so bili proteini, povezani s signalizacijo, membranskimi procesi, odzivom na stres, proteinsko homeostazo in metabolizmom lipidov. Neposredne vezave T3 na posamezne identificirane proteine v okviru te raziskave nismo eksperimentalno potrdili. Rezultati zato predstavljajo nabor perspektivnih kandidatov za nadaljnje raziskave in pomemben prvi korak k razumevanju možne vloge ščitničnim hormonom podobnih spojin ter njihovih potencialnih tarčnih proteinov v rastlinah. Triiodothyronine (T3) and thyroxine, also known as tetraiodothyronine (T4), play important roles in the regulation of development, growth, and metabolism in humans and other animals. In recent years, increasing evidence has suggested that thyroid hormone‑like compounds are also present in plants. However, their physiological roles and molecular mechanisms of action remain poorly understood. The aim of this master's thesis was to identify potential T3-binding proteins in the model plant Arabidopsis thaliana. To achieve this, T3 was immobilized on magnetic beads and used in pull down experiments with A. thaliana lysate. Elution fractions obtained by affinity chromatography were analyzed by mass spectrometry, and the identified proteins were functionally classified based on annotations from the UniProt database. In addition, fluorescently labeled T3 was prepared using Alexa Fluor 488 dye, incubated with plant lysate, and the interaction of the labeled hormone with plant proteins was analyzed by native gel electrophoresis. Mass spectrometry analysis enabled the identification of more than a hundred proteins, among which eight of the most robust candidates were highlighted using a combination of different statistical approaches. These proteins were associated with signaling, membrane-related processes, stress responses, protein homeostasis, and lipid metabolism. Direct binding of T3 to the identified proteins was not experimentally confirmed within the scope of this study. Nevertheless, the results provide a set of promising candidates for future investigations and represent an important first step toward understanding the possible role of thyroid hormone-like compounds and their potential target proteins in plants. test »pull down« Arabidopsis thaliana jod masna spektrometrija trijodotironin (T3) pull down assay Arabidopsis thaliana iodine mass spectrometry triiodothyronine (T3) true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-09-16 09:55:01 2026-09-16 09:55:08 2026-09-17 04:30:02 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 25543 Spruk_Teja_-_Identifikacija_vezavnih_partnerjev_rastlinskega_analoga_hormona_T3_med_proteini_nav.pdf Spruk_Teja_-_Identifikacija_vezavnih_partnerjev_rastlinskega_analoga_hormona_T3_med_proteini_nav.pdf 1 4DA87E0B91C757B8BBA6845D297CA53C b66cc56a920703a757b36e850ebfea484ec6ad3ccc9f5ae96310b2d2c1fd6c49 e6d528bf-b1a3-11f1-8bc5-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=247764 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0