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Substance P in the Gas Phase: Conformational Changes and Dissociations Induced by Collisional Activation in a Drift Tube

  • Christopher R. Conant
  • Daniel R. Fuller
  • Zhichao Zhang
  • Daniel W. Woodall
  • David H. Russell
  • David E. ClemmerEmail author
Focus: Ion Mobility Spectrometry (IMS): Research Article

Abstract

The work presented below is related to our companion paper in this issue, entitled: Substance P in solution: trans-to-cis configurational changes of penultimate prolines initiate non-enzymatic peptide bond cleavages. Two-dimensional ion mobility spectrometry (IMS-IMS) and mass spectrometry techniques are used to investigate structural transitions for [M+3H]3+ ions of substance P (subP) upon collisional activation (CA) in the gas phase. In this approach, different conformations of ions having a specified mobility are selected after an initial IMS separation, collisionally activated to produce new conformers, and these product structures are separated again using a second IMS region. In this way, it is possible to follow folding and unfolding transitions of different conformations. The analysis shows evidence for five conformations. Unlike other systems, every transition is irreversible. Studies as a function of activation voltage are used to discern pathways of structural changes prior to reaching the energy required for dissociation. Thresholds associated with the onsets of transitions are calibrated to obtain estimates of the energetic barriers between different structures and semi-quantitative potential energy diagrams are presented. Overall, barriers associated with structural transitions of [subP+3H]3+ in the absence of solvent are on the order of ~ 40 kJ mol−1, substantially lower than the ~ 90 kJ mol−1 required for some similar structural transitions in solutions of ethanol. Comparisons of the transition energies in the gas phase with thermochemistry for similar transitions in solution provide clues about why reverse transitions are prohibited.

Graphical Abstract

Keywords

Ion mobility spectrometry-mass spectrometry Peptide conformation Activation energy 

Notes

Acknowledgements

This work is supported in part by funds from the National Institute of Health, R01 GM117207-04, and the Indiana University Robert and Marjorie Mann fellowships (CRC, DRF). The work at TAMU (DHR) was funded by NSF (CHE-1707675) and NIH (P41GM121751-01A1).

Supplementary material

13361_2019_2160_MOESM1_ESM.docx (315 kb)
ESM 1 (DOCX 314 kb)

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© American Society for Mass Spectrometry 2019

Authors and Affiliations

  1. 1.Department of ChemistryIndiana UniversityBloomingtonUSA
  2. 2.Department of ChemistryTexas A&M UniversityCollege StationUSA

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