Interactions of Ligand, Aptamer, and Complementary Oligonucleotide: Studying Impacts of Na+ and Mg2+ Cations on Sensitive FRET-Based Detection of Aflatoxin B1

Samokhvalov, A.V.; Maksimenko, O.G.; Eremin, S.A.; Zherdev, A.V.; Dzantiev, B.B.

Авторы: Samokhvalov Alexey V., Maksimenko Oksana G., Eremin Sergei A., Zherdev Anatoly V., Dzantiev Boris B.
Журнал: Molecules
Том: 30
Номер: 10
Год издания: 2025
Издательство: MDPI
Местоположение издательства: Basel, Switzerland
Первая страница: 2125
Последняя страница: 2125
DOI: 10.3390/molecules30102125
Аннотация: The effects of magnesium and sodium on the interactions between aptamer, itsspecific ligand, and short complementary oligonucleotides (cDNAs) differing in affinityof their binding with the aptamer were studied. Aflatoxin B1 (AFB1) and AFB1-bindingaptamerwere used in the study. Dependencies for the aptamer bindingwith the fluorophorelabeledAFB1 under varied concentrations of the cations were obtained using fluorescenceanisotropy measurements. The increase of the aptamer affinity to AFB1 in the presence ofcations was demonstrated using fluorescence anisotropy and isothermal calorimetry. Thecollected data indicate that 300 mM Mg2+ (significantly more than the range commonlyused in aptamer sensors) provides the best affinity (16.5 ± 2.2 nM) of the aptamer–AFB1complexation. Sodium decreases the Mg2+-modulated affinity at some Na+/Mg2+ ratios.The aptamer affinity with cDNAs increases with concentration of cations, but not in the sameway as for AFB1. Based on the characterized conditions for bimolecular interactions, theligand-induced displacement of cDNAs was studied with the registration of the Forster fluorescenceenergy transfer (FRET). The most sensitive revealing of AFB1 (IC10% 3.2 ± 0.3 nM)in this three-compound FRET system was demonstrated for cDNA having an equilibriumconstant of the aptamer binding close to the constant of the aptamer–AFB1 reaction.
Добавил в систему: Еремин Сергей Александрович

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Interactions of Ligand, Aptamer, and Complementary Oligonucleotide: Studying Impacts of Na+ and Mg2+ Cations on Sensitive FRET-Based Detection of Aflatoxin B1статья Исследовательская статья

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