Sizing up DNA nanostructure assembly with native mass spectrometry and ion mobility

Jeroen F. Dyck, Jonathan R. Burns, Kyle I. P. Huray, Albert Konijnenberg, Stefan Howorka () and Frank Sobott ()
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Jeroen F. Dyck: University of Antwerp
Jonathan R. Burns: University College London
Kyle I. P. Huray: University of Leeds
Albert Konijnenberg: University of Antwerp
Stefan Howorka: University College London
Frank Sobott: University of Antwerp

Nature Communications, 2022, vol. 13, issue 1, 1-8

Abstract: Abstract Recent interest in biological and synthetic DNA nanostructures has highlighted the need for methods to comprehensively characterize intermediates and end products of multimeric DNA assembly. Here we use native mass spectrometry in combination with ion mobility to determine the mass, charge state and collision cross section of noncovalent DNA assemblies, and thereby elucidate their structural composition, oligomeric state, overall size and shape. We showcase the approach with a prototypical six-subunit DNA nanostructure to reveal how its assembly is governed by the ionic strength of the buffer, as well as how the mass and mobility of heterogeneous species can be well resolved by careful tuning of instrumental parameters. We find that the assembly of the hexameric, barrel-shaped complex is guided by positive cooperativity, while previously undetected higher-order 12- and 18-mer assemblies are assigned to defined larger-diameter geometric structures. Guided by our insight, ion mobility-mass spectrometry is poised to make significant contributions to understanding the formation and structural diversity of natural and synthetic oligonucleotide assemblies relevant in science and technology.

Date: 2022
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DOI: 10.1038/s41467-022-31029-5

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