Probing the Stability of a β-Hairpin Scaffold after Desolvation

Abstract

Probing the structural characteristics of biomolecular ions in the gas phase following native mass spectrometry (nMS) is of great interest, because noncovalent interactions, and thus native fold features, are believed to be largely retained upon desolvation. However, the conformation usually depends heavily on the charge state of the species investigated. In this study, we combine transition metal ion Forster resonance energy transfer (tmFRET) and ion mobility-mass spectrometry (IM-MS) with molecular dynamics (MD) simulations to interrogate the β-hairpin structure of GB1p in vacuo. Fluorescence lifetime values and collisional cross sections suggest an unfolding of the β-hairpin motif for higher charge states. MD simulations are consistent with experimental constraints, yet intriguingly provide an alternative structural interpretation: preservation of the β-hairpin is not only predicted for 2+ but also for 4+ charged species, which is unexpected given the substantial Coulomb repulsion for small secondary structure scaffolds.

Insights into conformational ensembles of compositionally identical disordered peptidomimetics

“…IMS and LC-IMS-MS are performed in non-aqueous environments (e.g., gas and organic phase), conformations of aqueous material have been demonstrated to be retained after desolvation for ion mobility using simulations 78,80 and we offer LC as an additional tool to capture these trends. As each of the selected model sequences contain the same composition, we hypothesize that conformational differences are maintained in these different environments.…”