Combining ion mobility spectrometry with hydrogen-deuterium exchange and top-down MS for peptide ion structure analysis

J Am Soc Mass Spectrom. 2014 Dec;25(12):2103-15. doi: 10.1007/s13361-014-0990-0. Epub 2014 Sep 30.

Abstract

The gas-phase conformations of electrosprayed ions of the model peptide KKDDDDIIKIIK have been examined by ion mobility spectrometry (IMS) and hydrogen deuterium exchange (HDX)-tandem mass spectrometry (MS/MS) techniques. [M+4H](4+) ions exhibit two conformers with collision cross sections of 418 Å(2) and 471 Å(2). [M+3H](3+) ions exhibit a predominant conformer with a collision cross section of 340 Å(2) as well as an unresolved conformer (shoulder) with a collision cross section of ~367 Å(2). Maximum HDX levels for the more compact [M+4H](4+) ions and the compact and partially-folded [M+3H](3+) ions are ~12.9, ~15.5, and ~14.9, respectively. Ion structures obtained from molecular dynamics simulations (MDS) suggest that this ordering of HDX level results from increased charge-site/exchange-site density for the more compact ions of lower charge. Additionally, a new model that includes two distance calculations (charge site to carbonyl group and carbonyl group to exchange site) for the computer-generated structures is shown to better correlate to the experimentally determined per-residue deuterium uptake. Future comparisons of IMS-HDX-MS data with structures obtained from MDS are discussed with respect to novel experiments that will reveal the HDX rates of individual residues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Deuterium Exchange Measurement / methods*
  • Ions / analysis
  • Ions / chemistry*
  • Mass Spectrometry / methods*
  • Molecular Dynamics Simulation
  • Peptides / analysis
  • Peptides / chemistry*
  • Protein Conformation

Substances

  • Ions
  • Peptides