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Charge site manipulation to enhance top-down fragmentation efficiency.

Tanja Habeck1, Edvaldo Vasconcelos Soares Maciel1, Kevin Kretschmer1

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|April 12, 2023
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Summary

Adding high-boiling solvents like DMSO to top-down mass spectrometry improves proteoform analysis. This method enhances fragmentation efficiency and sequence coverage for proteins such as alpha-synuclein.

Keywords:
electrospray ionization mass spectrometry (ESI-MS)mass spectrometrytop-down proteomics

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Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Top-down mass spectrometry is crucial for proteoform analysis.
  • Improving sequence coverage in top-down mass spectrometry remains a key challenge.

Purpose of the Study:

  • To investigate the effect of high-boiling solvents on top-down fragmentation patterns.
  • To determine if solvent additives can enhance sequence coverage and fragmentation efficiency.

Main Methods:

  • Proteins (alpha-synuclein, bovine carbonic anhydrase) were analyzed using top-down collision-induced dissociation (CID) mass spectrometry.
  • Low concentrations (2%) of DMSO and propylene carbonate were added to protein solutions.
  • Experiments were performed in triplicate to ensure reproducibility.

Main Results:

  • Addition of solvents reproducibly altered top-down CID fragmentation patterns.
  • Differences were attributed to solution condition-dependent selective formation of protonation site isomers.
  • Sequence coverage increased, with combined datasets showing up to 35% improvement in cleavage coverage.

Conclusions:

  • High-boiling solvents can enhance top-down mass spectrometry efficiency.
  • This approach offers a promising strategy to increase proteoform sequence coverage.
  • The method provides a simple yet effective way to improve proteomic analysis.