Zaviranje homodimerizacije proteaze Mpro in bakterijske histidin kinaze EnvZ z malomolekulskimi inhibitorji v bakteriji Escherichia coli

Starc, Gaja (avtor), Novinec, Marko (mentor)

Homodimerizacija proteinov je ključna za uravnavanje številnih procesov v celicah. Encimi morajo za katalitično aktivnost pogosto dimerizirati, na ta način je torej opravljanje morfološke funkcije uravnavano z oligomernim stanjem. S porastom pojava odpornosti na antibiotike in pojavom novih virusov, se vse več raziskav ukvarja tudi s preučevanjem dimerizacijske površine kot potencialne tarče v razvoju novih zdravil, Iskanje novih inhibitorjev dimerizacije pa terja učinkovite metode presejanja knjižnic malih molekul, ki omogočajo identifikacijo oligomernega stanja. Tekom diplomskega dela smo skušali preveriti uporabnost sistemov LEXGFP in TOXGFP za detekcijo homodimerizacije v bakteriji Escherichia coli pri presejanju knjižnic malomolekulskih inhibitorjev dimerizacije v celicah. Analizirali smo oligomerno stanje štirih ključnih bakterijskih oz. virusnih encimov, za katere so poznani inhibitorji dimerizacije – proteaze Mpro iz SARS-CoV-2, bakterijske histidin kinaze EnvZ iz E. coli, proteaze citomegalovirusa in proteaze herpes virusa povezanega s Kaposijevim sarkomom. V bakteriji Escherichia coli smo uspeli potrdili le homodimerizacijo prvih dveh, nato pa smo njuno oligomerno stanje analizirali še po uporabi malomolekulskih inhibitorjev homodimerizacije. Statistično značilne rezultate smo uspeli pridobiti le s sistemom TOXGFP, za potrditev uporabnosti tega sistema pri presejanju knjižnic inhibitorjev dimerizacije pa so potrebne dodatne raziskave.

• Jezik: Slovenski jezik
• Ključne besede: homodimerizacija, malomolekulski inhibitor dimerizacije, proteaza Mpro, bakterijska histidin kinaza EnvZ
• Vrsta gradiva: Diplomsko delo/naloga
• Tipologija: 2.11 - Diplomsko delo
• Organizacija: FKKT - Fakulteta za kemijo in kemijsko tehnologijo
• Leto izida: 2024
• PID: 20.500.12556/RUL-159158
• COBISS.SI-ID: 202796803
• Datum objave v RUL: 02.07.2024

STARC, Gaja, 2024, Zaviranje homodimerizacije proteaze Mpro in bakterijske histidin kinaze EnvZ z malomolekulskimi inhibitorji v bakteriji Escherichia coli [na spletu]. Diplomsko delo. [Dostopano 8 april 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=159158

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Inhibition of homodimerization of the Mpro protease and bacterial histidine kinase EnvZ by small molecule inhibitors in the bacterium Escherichia coli

Izvleček

Homodimerization is crucial for the regulation of numerous cellular processes. Enzymes often require dimerization for catalytic activity, thus fulfilling morphological functions is regulated by their oligomeric state. With the rise of antibiotic resistance and the emergence of new viruses, more research is focused on utilizing the dimerization surface as a potential target in drug development. Identifying novel dimerization inhibitors requires effective methods for screening libraries of small molecules, that allow the identification of the oligomeric state. With this thesis, we attempted to verify the potential of two systems for detecting homodimerization in Escherichia coli – namely LEXGFP and TOXGFP – as tolls for screening libraries of small molecule dimerization inhibitors in bacterial cells. We analysed the oligomeric state of four key bacterial or viral enzymes, for which dimerization inhibitors are known – Mpro protease from SARS-CoV-2, bacterial histidine kinase EnvZ from Escherichia coli, the cytomegalovirus protease, and the protease of the herpesvirus associated with Kaposi's sarcoma. We managed to confirm only the homodimerization of the first two. We then analysed their oligomeric state after the use of dimerization inhibitors. Statistically significant results were obtained only with the TOXGFP system, and further research is needed to confirm the usability of this system for screening libraries of dimerization inhibitors.

• Ključne besede: homodimerization, small molecule inhibitor, Mpro protease, bacterial histidine kinase EnvZ