4.8 Article

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

Journal

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 60, Issue 28, Pages 15390-15398

Publisher

WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202016757

Keywords

G-quadruplexes; native mass spectrometry; oligonucleotides; temperature-controlled mass spectrometry; thermodynamics

Funding

  1. Swiss National Science Foundation [200020_178765]
  2. Swiss National Science Foundation (SNF) [200020_178765] Funding Source: Swiss National Science Foundation (SNF)

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The study introduces a novel method using native mass spectrometry and a temperature-controlled nanoelectrospray ionization source to investigate interactions between proximal DNA domains. Results show changes in T-m and thermodynamic characteristics of proximal DNA domains depending on the number of domains, their position, and order in a single experiment.
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 T-m and thermodynamic characteristics of proximal DNA domains depending on the number of domains, their position, and order in a single experiment.

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