Karakterizacija vpliva aminokislinskih derivatov sukcinimida na aktivnost katepsina V

Hribar, Urban (author), Novinec, Marko (mentor)

Katepsin V spada med katepsinu L podobne katepsine iz družine papainu podobnih peptidaz. Je cisteinska lizosomska endopeptidaza, ki jo najdemo le pri višjih primatih. Izraža se v specifičnih tkivih, predvsem v priželjcu, kjer ima pomembno vlogo pri predstavitvi antigenov. Pri diplomski nalogi smo pregledali knjižnico spojin, ki so bile sintetizirane kot potencialni alosterični inhibitorji katepsinov K in S, in med njimi želeli poiskati alosterične inhibitorje katepsina V. Inhibicijske lastnosti smo določili iz kinetičnih meritev, narejenih z uporabo fluorimetrije. Znotraj knjižnice spojin na žalost nismo uspeli najti nobenega dobrega alosteričnega inhibitorja. Tri najbolj obetavne spojine smo uporabili za titracijo katepsina V, pri čemer smo ugotovili, da ti najverjetneje ne kažejo hiperbolične inhibicije, ki je bila opažena pri alosteričnih inhibitorjih katepsina K, in najverjetneje nimajo alosteričnega mehanizma. Te tri spojine smo tudi računalniško umestili na alosterično in aktivno mesto katepsina V s pomočjo algoritma Autodock Vina 1.1.2. Prav tako smo izmerili inhibicijo encimske razgradnje elastina za eno izmed treh spojin.

• Language: Slovenian
• Keywords: katepsin V, alosterična inhibicija, encimska kinetika, računalniško prileganje spojin, makromolekularni substrat.
• Work type: Bachelor thesis/paper
• Typology: 2.11 - Undergraduate Thesis
• Organization: FKKT - Faculty of Chemistry and Chemical Technology
• Year: 2019
• PID: 20.500.12556/RUL-109860
• COBISS.SI-ID: 1538391747
• Publication date in RUL: 09.09.2019

HRIBAR, Urban, 2019, Karakterizacija vpliva aminokislinskih derivatov sukcinimida na aktivnost katepsina V [online]. Bachelor’s thesis. [Accessed 15 April 2025]. Retrieved from: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=eng&id=109860

Secondary language

• Language: English

Abstract

Cathepsin V is a cathepsin L-like peptidase from the papain-like cysteine peptidase family. It is a lysosomal peptidase found only in higher primates. It is expressed only in specific tissues, primarily in the thymus where it plays a major role in antigen presentation. Our goal for this assignment was to identify allosteric inhibitors from a library of compounds designed as alosteric inhibitors of cathepsins K and S. The inhibitory properties of the compounds were inferred from the results of enzyme kinetic measurements taken using fluorometry. Unfortunately, we managed to find no good allosteric inhibitors from the library. We used the three most promising inhibitor compounds for titration of cathepsin V, which did not act as hyperbolic inhibitors as observed for allosteric inhibitors of cathepsin K, and are therefore likely not allosteric. Using the molecular docking algorithm Autodock Vina we docked all three compounds into the active site and allosteric site of cathepsin V. We also measured the inhibitory activity of one compound on the degradation of soluble elastin.

• Keywords: cathepsin V, alosteric inhibition, enzyme kinetics, computer docking, macromolecular substrate.