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

Autor: Ciara K O' Sullivan, Veronica Esposito, Miriam Jauset-Rubio, Vasso Skouridou, Markéta Svobodová, Teresa Mairal, Antonella Virgilio, Aldo Galeone
Přispěvatelé: O'Sullivan, C. K., Mairal, T., Jauset-Rubio, M., Svobodova, M., Skouridou, V., Esposito, V., Virgilio, A., Galeone, A.
Jazyk: angličtina
Rok vydání: 2021
Předmět:
0301 basic medicine
Circular dichroism
SPR
melting curves
01 natural sciences
lcsh:Chemistry
immune system diseases
Surface plasmon resonance
Polyacrylamide gel electrophoresis
lcsh:QH301-705.5
Spectroscopy
Molecular Structure
G-quadruplex
Chemistry
Seed Storage Proteins
General Medicine
Aptamers
Nucleotide
respiratory system
11-mer aptamer
Computer Science Applications
CD
BLI
Soybean Proteins
Electrospray ionization
Aptamer
β-conglutin
010402 general chemistry
Mass spectrometry
Catalysis
Article
Inorganic Chemistry
03 medical and health sciences
otorhinolaryngologic diseases
Physical and Theoretical Chemistry
Molecular Biology
MST
Microscale thermophoresis
G-quadruplex
11-mer aptamer
β-conglutin
PAGE
melting curves
CD
NMR
BLI
MST
SPR
Organic Chemistry
Globulins
Allergens
Antigens
Plant
NMR
0104 chemical sciences
respiratory tract diseases
PAGE
G-Quadruplexes
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
Biophysics
Nucleic Acid Conformation
Zdroj: International Journal of Molecular Sciences
Volume 22
Issue 3
International Journal of Molecular Sciences, Vol 22, Iss 1150, p 1150 (2021)
ISSN: 1422-0067
DOI: 10.3390/ijms22031150
Popis: In previous work, a 93-mer aptamer was selected against the anaphylactic allergen, &beta
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&prime
or 3&prime
end or at both ends. The TT-tail at the 5&prime
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.
Databáze: OpenAIRE
Externí odkaz: