Author, as appears in the article.: O’Sullivan CK; Mairal T; Jauset-Rubio M; Svobodova M; Skouridou V; Esposito V; Virgilio A; Galeone A

Department: Enginyeria Química

URV's Author/s: Jauset Rubio, Miriam / Mairal Lerga, Teresa / O'SULLIVAN, CIARA KATHLEEN / Skouridou, Vasoula / SVOBODOVÁ, MARKÉTA

Keywords: Β-conglutin Spr Page Nmr Mst Melting curves G-quadruplex Cd Bli 11-mer aptamer β -conglutin

Abstract: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. 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.

Thematic Areas: Zootecnia / recursos pesqueiros Spectroscopy Saúde coletiva Química Psicología Physical and theoretical chemistry Organic chemistry Odontología Nutrição Molecular biology Medicine (miscellaneous) Medicina veterinaria Medicina iii Medicina ii Medicina i Materiais Interdisciplinar Inorganic chemistry Geociências Farmacia Engenharias iv Engenharias ii Engenharias i Educação física Computer science applications Ciências biológicas iii Ciências biológicas ii Ciências biológicas i Ciências ambientais Ciências agrárias i Ciência de alimentos Ciência da computação Chemistry, multidisciplinary Catalysis Biotecnología Biodiversidade Biochemistry & molecular biology Astronomia / física

licence for use: https://creativecommons.org/licenses/by/3.0/es/

Author's mail: miriam.jauset@urv.cat vasoula.skouridou@urv.cat

Author identifier: 0000-0002-9943-6132 0000-0002-9712-5429

Record's date: 2023-02-23

Journal volume: 22

Papper version: info:eu-repo/semantics/publishedVersion

Link to the original source: https://www.mdpi.com/1422-0067/22/3/1150

Licence document URL: http://repositori.urv.cat/ca/proteccio-de-dades/

Papper original source: International Journal Of Molecular Sciences. 22 (3): 1-15

APA: O’Sullivan CK; Mairal T; Jauset-Rubio M; Svobodova M; Skouridou V; Esposito V; Virgilio A; Galeone A (2021). Aptamers against the β-conglutin allergen: Insights into the behavior of the shortest multimeric (intra)molecular dna gquadruplex. International Journal Of Molecular Sciences, 22(3), 1-15. DOI: 10.3390/ijms22031150

Article's DOI: 10.3390/ijms22031150

Entity: Universitat Rovira i Virgili

Journal publication year: 2021

Publication Type: Journal Publications