Compaction of Duplex Nucleic Acids upon Native ElectrosprayMass Spectrometry

摘要

We report on the fate of nucleic acids conformation in the gas phase as sampled using native mass spectrometry coupled to ion mobility spectrometry. On the basis of several successful reports for proteins and their complexes, the technique has become popular in structural biology, and the conformation survival becomes more and more taken for granted. Surprisingly, we found that DNA and RNA duplexes, at the electrospray charge states naturally obtained from native solution conditions (≥100 mM aqueous NH4OAc), are significantly more compact in the gas phase compared to the canonical solution structures. The compaction is observed for all duplex sizes (gas-phase structures are more compact than canonical B-helices by ∼20% for 12-bp, and by up to ∼30% for 36-bp duplexes), and for DNA and RNA alike. Molecular modeling (density functional calculations on small helices, semiempirical calculations on up to 12-bp, and molecular dynamics on up to 36-bp duplexes) demonstrates that the compaction is due to phosphate group self-solvation prevailing over Coulomb repulsion.Molecular dynamics simulations starting from solution structures donot reproduce the experimental compaction. To be experimentally relevant,molecular dynamics sampling should reflect the progressive structuralrearrangements occurring during desolvation. For nucleic acid duplexes,the compaction observed for low charge states results from novel phosphate–phosphatehydrogen bonds formed across both grooves at the very late stagesof electrospray.