期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 60, 期 28, 页码 15390-15398出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202016757
关键词
G-quadruplexes; native mass spectrometry; oligonucleotides; temperature-controlled mass spectrometry; thermodynamics
资金
- Swiss National Science Foundation [200020_178765]
- Swiss National Science Foundation (SNF) [200020_178765] Funding Source: Swiss National Science Foundation (SNF)
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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