diplomsko delo
Nika Mikulič Vernik (Avtor), Marko Novinec (Mentor)

Povzetek

Odkrivanje spojin s protimikrobnimi lastnostmi je odločilnega pomena za preprečevanje rastoče odpornosti proti antibiotikom. Za iskanje zdravil farmacevtska industrija uporablja pristop iskanja ligandov, ki vežejo (in s tem inhibirajo) tarčni protein. Da bi zaznali tudi interakcije z drugimi celičnimi komponentami in se s tem izognili spojinam s potencialno toksičnim vplivom, v našem laboratoriju uporabljamo pristop iskanja proteinskih tarč malih molekul. V diplomskem delu smo želeli določiti proteinske tarče male molekule 4-(2-aminoetil)-1-(piridin-2-il)-1H-pirzol-5-ol, za katero smo ugotovili, da zavira rast laboratorijskega seva DH5α bakterije Escherichia coli. Spojino smo imobilizirali na agarozni nosilec in pripravljeno kolono uporabili za afinitetno kromatografijo, pri kateri smo iz lizata E. coli izolirali interagirajoče proteine. Eluirane frakcije smo nato analizirali z NaDS-PAGE in poslali na analizo z masno spektrometrijo. Ugotovili smo, da dve intenzivni lisi na gelu predstavljata treonin dehidrogenazo in Fe–S podenoto sukcinat dehidrogenaze. Glede na literaturo je možen mehanizem bakteriostatičnega delovanja tarčne spojine inhibicija treonin dehidrogenaze, ki tako ni sposobna sinteze signalnih molekul. Inhibicija sukcinat dehidrogenaze verjetno ne vpliva na rast bakterije E. coli.

Ključne besede

kemoterapevtiki;antibiotiki;malomolekulski ligandi;proteini;proteinske tarče;afinitetna kromatografija;masna spektrometrija;medcelična signalizacija;diplomska dela;

Podatki

Jezik: Slovenski jezik
Leto izida:
Tipologija: 2.11 - Diplomsko delo
Organizacija: UL FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik: [N. Mikulič Vernik]
UDK: 577.18(043.2)
COBISS: 27183363 Povezava se bo odprla v novem oknu
Št. ogledov: 477
Št. prenosov: 165
Ocena: 0 (0 glasov)
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Ostali podatki

Sekundarni jezik: Angleški jezik
Sekundarni naslov: Identification of protein targets of a pyrazole derivative in Escherichia coli
Sekundarni povzetek: The discovery of compounds with antimicrobial properties is crucial to prevent the growing resistance to antibiotics. To search for drugs, the pharmaceutical industry utilises the approach of finding ligands that bind (and thus inhibit) the target protein. In order to detect interactions with other cellular components and thus avoid compounds with potentially toxic side effects, we used a small molecule to protein targeting approach in our laboratory. In this thesis, we aimed to identify the protein targets of the small molecule 4-(2-aminoethyl)-1-(pyridin-2-yl)-1H-pyrzol-5-ol, which was shown to inhibit the growth of Escherichia coli laboratory strain DH5α. The compound was immobilized on agarose beads and the prepared column was used for affinity chromatography with which we isolated interacting proteins from the E. coli lysate. Eluted fractions were analyzed by SDS-PAGE and sent for protein identification by mass spectrometry. Two prominent bands on the gel were identified as threonine dehydrogenase and the Fe-S subunit of succinate dehydrogenase. According to the literature, the possible mechanism for bacteriostatic action of the compound is inhibition of threonine dehydrogenase which is involved in the biosynthesis of signalling molecules. Inhibition of succinate dehydrogenase is unlikely to affect E. coli growth.
Sekundarne ključne besede: chemotherapeutics;low molecular weight ligand;affinity chromatography;mass spectrometry;quorum sensing;
Vrsta dela (COBISS): Diplomsko delo/naloga
Študijski program: 1000371
Komentar na gradivo: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo
Strani: 34 str.
ID: 12010395