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Native Electrospray Mass Spectrometry of DNA G-Quadruplexes in Potassium Solution

Abstract : A commonly used electrolyte in electrospray mass spectrometry (ESI-MS) of biomolecules is ammonium acetate (NH 4 OAc). Although some nucleic acid structures such as duplexes require only proper physiological ionic strength (whatever the monovalent ions) to be properly folded in ESI-MS conditions, the folding of some other nucleic acid structures such as DNA G-quadruplexes also depends on direct binding of specific cations. Here, we developed ESI-MS compatible conditions that allow one to observe DNA G-quaduplexes with K + ions specifically bound between G-quartets. NH 4 OAc was replaced with trimethylammonium acetate (TMAA), at concentrations up to 150 mM to provide physiological ionic strength, and the solution was doped with KCl at concentrations up to 1 mM. The trimethylammonium ion is too large to coordinate between G-quartets, where only K + ions bind. Compared with the equivalent NH 4 OAc/KCl mixtures, the TMAA/KCl mixtures provide cleaner spectra by suppressing the nonspecific adducts, and favor the formation of similar stacking arrangements as in 100 mM KCl (physiologically relevant cation) for the polymorphic human telomeric DNA G-quadruplexes. This new sample preparation method can be exploited to determine the number of potassium binding sites in new sequences, to screen ligand binding to the structures favored in potassium, and to transfer potassium-bound G-quadruplexes to the mass spectrometer for gas-phase structural probing, as illustrated herein with ion mobility spectrometry experiments.
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Contributor : Valérie Gabelica Connect in order to contact the contributor
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2014-JASMS_G4inK_TMAA (1).pdf
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Adrien Marchand, Valérie Gabelica. Native Electrospray Mass Spectrometry of DNA G-Quadruplexes in Potassium Solution. Journal of The American Society for Mass Spectrometry, American Chemical Society 2014, 25 (7), pp.1146. ⟨10.1007/s13361-014-0890-3⟩. ⟨inserm-01385783⟩



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