Istraživanje interakcija cijaninskih boja s polinukleotidima spektroskopskim i termoanalitičkim metodama

doctoral thesis

• Title (english): Study of cyanine dyes interactions with polynucleotides using spectroscopic and thermoanalytical methods of analyses
• Author: Andrea Rožman
• Mentor: Ivo Piantanida (mentor)
• Committee member: Vladislav Tomišić (predsjednik povjerenstva)
• Committee member: Snežana Miljanić (član povjerenstva)
• Committee member: Ivo Piantanida (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2018-12-13, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry
• Universal decimal classification (UDC): 54 - Chemistry. Crystallography. Mineralogy

Abstract

Uvođenje klora kao substituenta na cijaninski kromofor dobro poznatih cijaninskih boja TOTO i
YOYO, koje se koriste za bojanje i vizualizaciju DNA, rezultirao je značajnim promjenama u
molekulskom prepoznavanju različitih DNA i RNA u usporedbi s ishodnim bojama, a isto tako i u
odnosu na odgovarajuće monomere 6-Cl-TO-1 i 6-Cl-YO-1.
Istraživani novi spojevi tvore nekovalentne komplekse s dvolančanim DNA/RNA, vrlo visoke
stabilnosti, Ks > 107 dm3 mol−1, s malim razlikama u konstantama stabilnosti reakcije vezanja između
različitih sljedova DNA/RNA. Spoj 6-Cl-YOYO-1 je pokazao specifično fluorimetrijsko
prepoznavanje ds-RNA karakterističnom emisijom na 570 nm, koja se ne pojavljuje dodatkom bilo
koje ds-DNA. Navedena specifična emisija (570 nm) pripisuje se svojstvu agregacije 6-Cl-YOYO-1
boje, te stoga nije prisutna kod analogne boje 6-Cl-TOTO-1. 6-Cl-TOTO-1 je za razliku od YO
analoga, pokazao vrpcu induciranog cirkularnog dikroizma (ICD) pri 520 nm samo prilikom vezanja
na alternirajuće DNA sekvence (GC-GC i AT-AT), što nije primijećeno prilikom vezanja na homo-
DNA i homo-RNA. Takav obrazac prepoznavanja pripisuje se strukturnim razlikama između
polinukleotida, gdje samo alternirajuće DNA imaju tipičnu B-helikalnu strukturu. Dakle, oba
klorirana analoga istraživanih boja pokazuju nova svojstva molekulskog prepoznavanja u odnosu na
ishodne TOTO i YOYO, koje u kombinaciji sa zanemarivom citotoksičnošću otvaraju potencijal za
razvoj novih boja za biokemijske i biomedicinske aplikacije.

Abstract (english)

Introduction of chloro-substituent at cyanine chromophore of well-known DNA stains TOTO and
YOYO resulted in a significant difference of DNA and RNA recognition with respect to parent
compounds (TOTO; YOYO), as well as in respect to corresponding monomers (6-Cl-TO-1 and 6-Cl-
YO-1). Studied novel compounds form non-covalent complexes with ds-DNA/RNA which exhibited
high stability Ks > 107 dm3 mol−1, with minor differences in binding constants between various DNA
and RNA. 6-Cl-YOYO-1 revealed specific fluorimetric recognition of ds-RNA in comparison to ds-
DNA by the fluorimetric response at 570 nm, which was attributed to particular aggregation property
of the dye, not present in corresponding 6-Cl-TOTO-1 analogue. The 6-Cl-TOTO-1, unlike YOanalogue,
showed ICD band at 520 nm only upon binding to alternating DNA sequences (both, GCGC
and AT-AT), which was not observed for homo-DNA and homo-RNA. Such recognition pattern
was attributed to structural differences between polynucleotides, whereby only alternating-DNAs have
a typical B-helical structure in which 6-Cl-TOTO-1 molecule could insert to give particular ICD band
at 520 nm. Thus, both Cl-analogues showed intriguing novel recognition properties in comparison to
parent TOTO and YOYO, which combined with negligible cytotoxicity offer potential for
development of novel dyes for biochemical and biomedical applications.

• Keywords: ciljana agregacija boja; cirkularni dikroizam; DNA i RNA prepoznavanje; fluorescentni markeri
• Keywords (english): circular dichroism; DNA and RNA recognition; fluorescent markers; target-aggregating dyes
• Language: croatian
• URN:NBN: urn:nbn:hr:217:721510
• Study programme: Title: Doctoral study in chemistry; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje prirodnih znanosti, polje kemija (doktor/doktorica znanosti, područje prirodnih znanosti, polje kemija)
• Type of resource: Text
• Extent: 143 str. ; 30 cm
• File origin: Born digital
• Access conditions: Open access
• Created on: 2019-01-08 10:28:24