Novel Insight into Proximal DNA Domain Interactions from Temperature‐Controlled Electrospray Ionization Mass Spectrometry

Quadruplexes are non‐canonical nucleic acid structures essential for many cellular processes. Hybrid quadruplex–duplex oligonucleotide assemblies comprised of multiple domains are challenging to study with conventional biophysical methods due to their structural complexity. Here, we introduce a nove...

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Bibliographic Details
Published in:Angewandte Chemie 2021-07, Vol.133 (28), p.15518-15526
Main Authors: Pruška, Adam, Marchand, Adrien, Zenobi, Renato
Format: Article
Language:English
Subjects:
Cellular structure
Chemistry
Deoxyribonucleic acid
DNA
Domains
G-quadruplexes
Intermediates
Ionization
Ions
Mass spectrometry
Mass spectroscopy
native mass spectrometry
Nucleic acids
Oligonucleotides
Scientific imaging
Spectroscopy
Temperature
temperature-controlled mass spectrometry
Thermal denaturation
thermodynamics
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Summary:Quadruplexes are non‐canonical nucleic acid structures essential for many cellular processes. Hybrid quadruplex–duplex oligonucleotide assemblies comprised of multiple domains are challenging to study with conventional biophysical methods due to their structural complexity. Here, we introduce a novel method based on native mass spectrometry (MS) coupled with a custom‐built temperature‐controlled nanoelectrospray ionization (TCnESI) source designed to investigate interactions between proximal DNA domains. Thermal denaturation experiments were aimed to study unfolding of multi‐stranded oligonucleotide constructs derived from biologically relevant structures and to identify unfolding intermediates. Using the TCnESI MS, we observed changes in Tm and thermodynamic characteristics of proximal DNA domains depending on the number of domains, their position, and order in a single experiment. Hybrid duplex–quadruplex oligonucleotides comprised of multiple domains have not been yet properly investigated because of their structural complexity. A new method based on native mass spectrometry (MS) coupled with a custom‐built temperature‐controlled nanoelectrospray ionization (TCnESI) source is introduced to investigate effects between multiple proximal DNA domains.
ISSN:0044-8249
1521-3757
DOI:10.1002/ange.202016757