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Open AccessArticle

Aptamers Against the β-Conglutin Allergen: Insights into the Behavior of the Shortest Multimeric(Intra)Molecular DNA G-Quadruplex

1
INTERFIBIO Research Group, Departament d’Enginyeria Química, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007 Tarragona, Spain
2
Institució Catalana de Recerca i Estudis Avançats (ICREA), Passeig Lluís Companys 23, 08010 Barcelona, Spain
3
Dipartimento di Farmacia, Università degli Studi di Napoli Federico II, Via D. Montesano 49, 80131 Napoli, Italy
*
Author to whom correspondence should be addressed.
These authors have contributed equally to the work.
Int. J. Mol. Sci. 2021, 22(3), 1150; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22031150
Received: 28 December 2020 / Revised: 13 January 2021 / Accepted: 19 January 2021 / Published: 24 January 2021
In previous work, a 93-mer aptamer was selected against the anaphylactic allergen, β-conglutin and truncated to an 11-mer, improving the affinity by two orders of magnitude, whilst maintaining the specificity. This 11-mer was observed to fold in a G-quadruplex, and preliminary results indicated the existence of a combination of monomeric and higher-order structures. Building on this previous work, in the current study, we aimed to elucidate a deeper understanding of the structural forms of this 11-mer and the effect of the structure on its binding ability. A battery of techniques including polyacrylamide gel electrophoresis, high-performance liquid chromatography in combination with electrospray ionization time-of-flight mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight, thermal binding analysis, circular dichroism and nuclear magnetic resonance were used to probe the structure of both the 11-mer and the 11-mer flanked with TT- at either the 5′ or 3′ end or at both ends. The TT-tail at the 5′ end hinders stacking effects and effectively enforces the 11-mer to maintain a monomeric form. The 11-mer and the TT- derivatives of the 11-mer were also evaluated for their ability to bind its cognate target using microscale thermophoresis and surface plasmon resonance, and biolayer interferometry confirmed the nanomolar affinity of the 11-mer. All the techniques utilized confirmed that the 11-mer was found to exist in a combination of monomeric and higher-order structures, and that independent of the structural form present, nanomolar affinity was observed. View Full-Text
Keywords: G-quadruplex; 11-mer aptamer; β-conglutin; PAGE; melting curves; CD; NMR; BLI; MST; SPR G-quadruplex; 11-mer aptamer; β-conglutin; PAGE; melting curves; CD; NMR; BLI; MST; SPR
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MDPI and ACS Style

O’ Sullivan, C.K.; Mairal, T.; Jauset-Rubio, M.; Svobodova, M.; Skouridou, V.; Esposito, V.; Virgilio, A.; Galeone, A. Aptamers Against the β-Conglutin Allergen: Insights into the Behavior of the Shortest Multimeric(Intra)Molecular DNA G-Quadruplex. Int. J. Mol. Sci. 2021, 22, 1150. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22031150

AMA Style

O’ Sullivan CK, Mairal T, Jauset-Rubio M, Svobodova M, Skouridou V, Esposito V, Virgilio A, Galeone A. Aptamers Against the β-Conglutin Allergen: Insights into the Behavior of the Shortest Multimeric(Intra)Molecular DNA G-Quadruplex. International Journal of Molecular Sciences. 2021; 22(3):1150. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22031150

Chicago/Turabian Style

O’ Sullivan, Ciara K.; Mairal, Teresa; Jauset-Rubio, Miriam; Svobodova, Marketa; Skouridou, Vasso; Esposito, Veronica; Virgilio, Antonella; Galeone, Aldo. 2021. "Aptamers Against the β-Conglutin Allergen: Insights into the Behavior of the Shortest Multimeric(Intra)Molecular DNA G-Quadruplex" Int. J. Mol. Sci. 22, no. 3: 1150. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22031150

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