Negative ion fragmentations of disulfide-containing cross-linking reagents are competitive with aspartic acid side-chain-induced cleavages

A N Calabrese; Tianfang Wang; J H Bowie; T L Pukala

doi:10.1002/rcm.6445

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Negative ion fragmentations of disulfide-containing cross-linking reagents are competitive with aspartic acid side-chain-induced cleavages

Journal article

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Negative ion fragmentations of disulfide-containing cross-linking reagents are competitive with aspartic acid side-chain-induced cleavages

A N Calabrese, Tianfang Wang, J H Bowie and T L Pukala

Rapid Communications in Mass Spectrometry, Vol.27(1), pp.238-248

2013

DOI: https://doi.org/10.1002/rcm.6445

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Abstract

RATIONALE: It has been shown that the disulfide moiety in the chemical cross-linking reagent dithiobis(succinimidyl)propionate (DSP), which is similar in structure to the natural cystine disulfide, cleaves preferentially to the peptide backbone in the negative ion mode. However, the tandem mass (MS/MS) spectra of peptides in the negative ion mode are often dominated by products arising from low-energy, side-chain-induced processes, which may compete with any facile cross-linker fragmentations and complicate identification of chemical cross-links in a complex mixture. METHODS: Two disulfide-containing crosslinking reagents similar to DSP, but with varying spacer arm lengths, were synthesized and the MS/MS spectra of several model peptides cross-linked with these reagents were investigated. Theoretical calculations were used to describe the energetics of the cross-linker fragmentations as well as several low-energy side-chain-induced fragmentations which compete with disulfide cleavages. RESULTS: Altering the spacer arm length of the cross-linker, such that there is one methylene group less than in DSP, results in a more facile cleavage process, whilst the opposite is true when a methylene group is added. Of the low-energy side-chain-induced fragmentations studied, only those from aspartic acid compete significantly with those of the cross-linker disulfide. CONCLUSIONS: Low-energy cleavage processes from aspartic acid that compete with cross-linker fragmentations occur in the negative ion MS/MS spectra of the cross-linked peptides, irrespective of the spacer arm length. Other fragmentation pathways do not significantly interfere with low-energy disulfide cleavage, making the presence of additional product ions in the MS/MS spectrum diagnostic for the presence of aspartic acid.

Details

Title: Negative ion fragmentations of disulfide-containing cross-linking reagents are competitive with aspartic acid side-chain-induced cleavages
Authors: A N Calabrese (Author) - University of Adelaide
Tianfang Wang (Author) - University of the Sunshine Coast - Faculty of Science, Health, Education and Engineering
J H Bowie (Author) - University of Adelaide
T L Pukala (Author) - University of Adelaide
Publication details: Rapid Communications in Mass Spectrometry, Vol.27(1), pp.238-248
Publisher: John Wiley & Sons Ltd.
Date published: 2013
DOI: 10.1002/rcm.6445
ISSN: 0951-4198
Organisation Unit: School of Science and Engineering - Legacy; University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering; Centre for Bioinnovation
Language: English
Record Identifier: 99450143702621
Output Type: Journal article

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Collaboration types: Domestic collaboration
Web Of Science research areas: Biochemical Research Methods; Chemistry, Analytical; Spectroscopy

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