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Evidence for Sequence Scrambling and Divergent H/D Exchange Reactions of Doubly-Charged Isobaric b-Type Fragment Ions

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

To date, only a limited number of reports are available on structural variants of multiply-charged b-fragment ions. We report on observed bimodal gas-phase hydrogen/deuterium exchange (HDX) reaction kinetics and patterns for substance P b10 2+ that point to presence of isomeric structures. We also compare HDX reactions, post-ion mobility/collision-induced dissociation (post-IM/CID), and sustained off-resonance irradiation-collision induced dissociation (SORI-CID) of substance P b10 2+ and a cyclic peptide with an identical amino acid (AA) sequence order to substance P b10. The observed HDX patterns and reaction kinetics and SORI-CID pattern for the doubly charged head-to-tail cyclized peptide were different from either of the presumed isomers of substance P b10 2+, suggesting that b10 2+ may not exist exclusively as a head-to-tail cyclized structure. Ultra-high mass measurement accuracy was used to assign identities of the observed SORI-CID fragment ions of substance P b10 2+; over 30 % of the observed SORI-CID fragment ions from substance P b10 2+ had rearranged (scrambled) AA sequences. Moreover, post-IM/CID experiments revealed the presence of two conformer types for substance P b10 2+, whereas only one conformer type was observed for the head-to-tail cyclized peptide. We also show that AA sequence scrambling from CID of doubly-charged b-fragment ions is not unique to substance P b10 2+.

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Acknowledgments

The authors gratefully acknowledge the financial support from the Institute for Therapeutic Discovery (Delanson, NY, USA) and Baylor University.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Baylor University, Waco, TX, 76798, USA

    Behrooz Zekavat, Mahsan Miladi & Touradj Solouki

  2. Department of Chemistry, University of Maine, Orono, ME, 04469-5706, USA

    Abdullah H. Al-Fdeilat

  3. Department of Chemistry and Biochemistry, University of Arizona, Tucson, AZ, 85721, USA

    Arpad Somogyi

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  1. Behrooz Zekavat
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  2. Mahsan Miladi
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  3. Abdullah H. Al-Fdeilat
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  4. Arpad Somogyi
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  5. Touradj Solouki
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Correspondence to Touradj Solouki.

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Zekavat, B., Miladi, M., Al-Fdeilat, A.H. et al. Evidence for Sequence Scrambling and Divergent H/D Exchange Reactions of Doubly-Charged Isobaric b-Type Fragment Ions. J. Am. Soc. Mass Spectrom. 25, 226–236 (2014). https://doi.org/10.1007/s13361-013-0768-9

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