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Article Dans Une Revue Analytical Chemistry Année : 2020

Native Hydrogen/Deuterium Exchange Mass Spectrometry of Struc- tured DNA Oligonucleotides

Résumé

Although solution hydrogen-deuterium exchange mass spectrometry (HDX/MS) is well-established for the analysis of the structure and dynamics of proteins, it is currently not exploited for nucleic acids. Here we used DNA G-quadruplex structures as model systems to demonstrate that DNA oligonucleotides are amenable to in-solution HDX/MS in native conditions. In trime-thylammonium acetate solutions and in soft source conditions, the protonated phosphate groups are fully back-exchanged in the source, while the exchanged nucleobases remain labeled without detectable back-exchange. As a result, the exchange rates depend strongly on the secondary structure (hydrogen bonding status) of the oligonucleotides, but neither on their charge state nor on the presence of non-specific adducts. We show that native mass spectrometry methods can measure these exchange rates on the second to the day time scale with high precision. Such combination of HDX with native MS opens promising avenues for the analysis of the structural and biophysical properties of oligonucleotides and their complexes.
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Dates et versions

hal-02882153 , version 1 (26-06-2020)

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Eric Largy, Valérie Gabelica. Native Hydrogen/Deuterium Exchange Mass Spectrometry of Struc- tured DNA Oligonucleotides. Analytical Chemistry, 2020, 92 (6), pp.4402-4410. ⟨10.1021/acs.analchem.9b05298⟩. ⟨hal-02882153⟩
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