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G-Quadruplex Aptamer-Ligand Characterization
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SYSNO ASEP 0563712 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název G-Quadruplex Aptamer-Ligand Characterization Tvůrce(i) Moreira, D. (PT)
Leitao, D. (PT)
Lopes-Nunes, J. (PT)
Santos, T. C. B. (PT)
Figueiredo, J. (PT)
Miranda, A.I. (PT)
Alexandre, D. (PT)
Tomaz, C. (PT)
Mergny, Jean-Louis (BFU-R) ORCID, RID
Cruz, C. (PT)Celkový počet autorů 10 Číslo článku 6781 Zdroj.dok. Molecules. - : MDPI
Roč. 27, č. 20 (2022)Poč.str. 16 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. CH - Švýcarsko Klíč. slova G-quadruplex aptamer ; ligands ; aptamer-ligand interactions ; biophysical techniques Vědní obor RIV CE - Biochemie Obor OECD Biochemistry and molecular biology CEP EF15_003/0000477 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Způsob publikování Open access Institucionální podpora BFU-R - RVO:68081707 UT WOS 000872819000001 EID SCOPUS 85140917571 DOI 10.3390/molecules27206781 Anotace In this work we explore the structure of a G-rich DNA aptamer termed AT11-L2 (TGGTGGTGGTTGTTGTTGGTGGTGGTGGT, derivative of AT11) by evaluating the formation and stability of G-quadruplex (G4) conformation under different experimental conditions such as KCl concentration, temperature, and upon binding with a variety of G4 ligands (360A, BRACO-19, PDS, PhenDC3, TMPyP4). We also determined whether nucleolin (NCL) can be a target of AT11-L2 G4. Firstly, we assessed by circular dichroism, UV and NMR spectroscopies the formation of G4 by AT11-L2. We observed that, for KCl concentrations of 65 mM or less, AT11-L2 adopts hybrid or multiple topologies. In contrast, a parallel topology predominates for buffer containing 100 mM of KCl. The T-m of AT11-L2 in 100 mM of KCl is 38.9 degrees C, proving the weak stability of this sequence. We also found that upon titration with two molar equivalents of 360A, BRACO-19 and PhenDC3, the G4 is strongly stabilized and its topology is maintained, while the addition of 3.5 molar equivalents of TMPyP4 promotes the disruption of G4. The K-D values between AT11-L2 G4, ligands and NCL were obtained by fluorescence titrations and are in the range of mu M for ligand complexes and nM when adding NCL. In silico studies suggest that four ligands bind to the AT11-L2 G4 structure by stacking interactions, while the RBD1,2 domains of NCL interact preferentially with the thymines of AT11-L2 G4. Finally, AT11-L2 G4 co-localized with NCL in NCL-positive tongue squamous cell carcinoma cell line. Pracoviště Biofyzikální ústav Kontakt Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244 Rok sběru 2023 Elektronická adresa https://www.mdpi.com/1420-3049/27/20/6781
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