Linear and Differential Ion Mobility Separations of Middle-Down Proteoforms
| Autor: | Ole N. Jensen, Todd D. Williams, Kevin Jeanne Dit Fouque, Alexandre A. Shvartsburg, Matthew A. Baird, Alyssa Garabedian, Jacob Porter, Pavel V. Shliaha, Francisco Fernandez-Lima |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Proteome Resolution (mass spectrometry) Tandem Chemistry Ion-mobility spectrometry Electrospray ionization 010401 analytical chemistry Transferability Mass spectrometry 01 natural sciences Mass spectrometric Article Mass Spectrometry 0104 chemical sciences Analytical Chemistry Ion Histones 03 medical and health sciences 030104 developmental biology Journal Article Peptides Biological system Protein Processing Post-Translational |
| Zdroj: | Garabedian, A, Baird, M A, Porter, J, Jeanne Dit Fouque, K, Shliaha, P V, Jensen, O N, Williams, T D, Fernandez-Lima, F & Shvartsburg, A A 2018, ' Linear and Differential Ion Mobility Separations of Middle-Down Proteoforms ', Analytical Chemistry, vol. 90, no. 4, pp. 2918-2925 . https://doi.org/10.1021/acs.analchem.7b05224 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.7b05224 |
| Popis: | Comprehensive characterization of proteomes comprising the same proteins with distinct post-translational modifications (PTMs) is a staggering challenge. Many such proteoforms are isomers (localization variants) that require separation followed by top-down or middle-down mass spectrometric analyses, but condensed-phase separations are ineffective in those size ranges. The variants for "middle-down" peptides were resolved by differential ion mobility spectrometry (FAIMS), relying on the mobility increment at high electric fields, but not previously by linear IMS on the basis of absolute mobility. We now use complete histone tails with diverse PTMs on alternative sites to demonstrate that high-resolution linear IMS, here trapped IMS (TIMS), broadly resolves the variants of ∼50 residues in full or into binary mixtures quantifiable by tandem MS, largely thanks to orthogonal separations across charge states. Separations using traveling-wave (TWIMS) and/or involving various time scales and electrospray ionization source conditions are similar (with lower resolution for TWIMS), showing the transferability of results across linear IMS instruments. The linear IMS and FAIMS dimensions are substantially orthogonal, suggesting FAIMS/IMS/MS as a powerful platform for proteoform analyses. |
| Databáze: | OpenAIRE |
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