Abstract
Generation of overlapping peptides in solution via multiple proteases requires a very high peak capacity for the LC-MS analysis to minimize signal overlap. An inherent advantage of a gas-phase fragmentation step is that the additional gas-phase fragment ions used to sublocalize deuterium are produced after precursor ion selection and thus do not add complexity to the LC-MS analysis. The key to obtaining optimal spatial resolution in a hydrogen exchange mass spectrometry (HX-MS) experiment is the fragmentation efficiency. This chapter discusses common fragmentation techniques like collision-induced dissociation (CID) occur with complete Hydrogen-deuterium (H/D) scrambling, while other techniques that induce dissociation on a faster timescale through radical-based fragmentation channels, like electron-capture dissociation (ECD) and electron-transfer dissociation (ETD), occur inherently without H/D scrambling, thus making them suitable for HX applications. By combining the classic bottom-up HX-MS workflow with gas-phase fragmentation by ETD, detailed information on protein HX can be obtained.
| Originalsprog | Engelsk |
|---|---|
| Titel | Hydrogen Exchange Mass Spectrometry of Proteins : Fundamentals, Methods and Applications |
| Redaktører | David Weis |
| Antal sider | 22 |
| Forlag | Wiley |
| Publikationsdato | mar. 2016 |
| Sider | 127-148 |
| Kapitel | 8 |
| ISBN (Trykt) | 978-1-118-61649-9 |
| Status | Udgivet - mar. 2016 |