Improved Protein and PTM Characterization with a Practical Electron-Based Fragmentation on Q-TOF Instruments
| Autor: | Valery G. Voinov, Michael Hare, George Stafford, Jared B. Shaw, Yury V. Vasil’ev, Derrill Sturgeon, Cody Schwarzer, Christian Klein, Rebecca Glaskin, Joseph S. Beckman, Shuai Wu, Diana Oppenheimer |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Phosphopeptides
Glycosylation Proline Electrons electron transfer dissociation Peptide Substance P 010402 general chemistry Proteomics 01 natural sciences Mass Spectrometry Isoaspartate chemistry.chemical_compound proteomics Leucine Structural Biology Insulin Disulfides electron capture dissociation Phosphorylation Fragmentation (cell biology) Spectroscopy chemistry.chemical_classification Isoaspartic Acid Electron-capture dissociation isoaspartate Lysine 010401 analytical chemistry Proteins proteoform 0104 chemical sciences Electron-transfer dissociation chemistry Biophysics Mass spectrum Protein Processing Post-Translational Software Research Article |
| Zdroj: | Journal of the American Society for Mass Spectrometry |
| ISSN: | 1879-1123 1044-0305 |
| DOI: | 10.1021/jasms.0c00482 |
| Popis: | Electron-based dissociation (ExD) produces uncluttered mass spectra of intact proteins while preserving labile post-translational modifications. However, technical challenges have limited this option to only a few high-end mass spectrometers. We have developed an efficient ExD cell that can be retrofitted in less than an hour into current LC/Q-TOF instruments. Supporting software has been developed to acquire, process, and annotate peptide and protein ExD fragmentation spectra. In addition to producing complementary fragmentation, ExD spectra enable many isobaric leucine/isoleucine and isoaspartate/aspartate pairs to be distinguished by side-chain fragmentation. The ExD cell preserves phosphorylation and glycosylation modifications. It also fragments longer peptides more efficiently to reveal signaling cross-talk between multiple post-translational modifications on the same protein chain and cleaves disulfide bonds in cystine knotted proteins and intact antibodies. The ability of the ExD cell to combine collisional activation with electron fragmentation enables more complete sequence coverage by disrupting intramolecular electrostatic interactions that can hold fragments of large peptides and proteins together. These enhanced capabilities made possible by the ExD cell expand the size of peptides and proteins that can be analyzed as well as the analytical certainty of characterizing their post-translational modifications. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|