Istraživanje interakcija klaritromicina i ribosoma izoliranog iz bakterije Escherichia coli spektroskopijom NMR

master's thesis

• Title (english): Study of interactions between clarithromycin and ribosome isolated from Escherichia coli by NMR spectroscopy
• Author: Mateja Tkalčević
• Mentor: Tomislav Jednačak (mentor)
• Committee member: Tomislav Jednačak (predsjednik povjerenstva)
• Committee member: Nikola Bregović (član povjerenstva)
• Committee member: Jasmina Rokov Plavec (član povjerenstva)
• Committee member: Ivana Biljan (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2021-09-28, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

Klaritromicin je predstavnik druge generacije makrolidnih antibiotika, u koju spada i poznatiji
azitromicin. Makrolidni antibiotici (makrolidi) se koriste u liječenju infekcija uzrokovanih
Gram-pozitivnim i Gram-negativnim bakterijama. Vezanjem za veliku podjedinicu
bakterijskog ribosoma makrolidi inhibiraju translaciju proteina čime ubijaju bakterijske stanice.
Učestalom primjenom makrolida bakterije su razvile različite mehanizme otpornosti na njihovo
djelovanje. Stoga je interes znanosti preusmjeren na razvoj novih makrolidnih antibiotika. Za
uspješan dizajn novog makrolida važno je razumjeti njegove interakcije s bakterijskim
ribosomom. Stoga je cilj ovog rada okarakterizirati strukturu i interakcije klaritromcina s
ribosomom izoliranim iz bakterije Escherichia coli u otopini primjenom jedno- i
dvodimenzijskih tehnika NMR. Tehnikama NOESY (engl. nuclear Overhauser effect
spectroscopy) i ROESY (engl. rotating frame Overhauser effect spectroscopy) određena je
konformacija klaritromicina u slobodnom stanju, a pomoću prijenosnog NOESY određena je
konformacija vezanog klaritromicina. Nadalje, metodom STD (engl. saturation transfer
difference) proučene su interakcije klaritromicina s ribosomom iz bakterije E. coli i određeni
su vezni epitopi.

Abstract (english)

Clarithromycin is a representative of the second generation of macrolide antibiotics, which
includes better-known azithromycin. Macrolides are used for treating infections caused by
Gram-positive and Gram-negative bacteria. Binding of macrolides to a large subunit of the
bacterial ribosome inhibits protein translation and kills bacterial cells. Due to a frequent use of
macrolides, bacteria have developed different resistance mechanisms. For this reason, the
interest of science has been redirected to the development of new macrolide antibiotics. For a
successful design of a new macrolide, it is important to understand its interactions with the
bacterial ribosome. Therefore, the purpose of this study is to characterize the structure and
interactions of clarithromycin with ribosome isolated from Escherichia coli in solution using
one- and two-dimensional NMR techniques. Conformational studies of clarithromycin in the
free-state were carried out using NOESY and ROESY techniques, while the bound-state
conformation was determined from trNOESY correlations. Furthermore, the interactions of
clarithromycin with the E. coli ribosome were studied by STD method, which allowed the
assessment of binding epitopes.

• Keywords: interakcije; klaritromicin; makrolidi; ribosom; spektroskopija NMR
• Keywords (english): clarithromycin; interactions; NMR spectroscopy; macrolides; ribosome
• Language: croatian
• URN:NBN: urn:nbn:hr:217:714511
• Project: Number: IP-2018-01-8098; Title: Makrozoni, novi konjugati makrolidnih antibiotika: Dizajn, priprava i interakcije; Title: Macrozones, New Conjugates of Macrolide Antibiotics: Design, Synthesis and Interactions; Acronym: Makrozoni/Macrozones; Leader: Predrag Novak; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-10-22 12:19:15