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Ion source parameters and hydrogen scrambling in the ECD of selectively deuterated peptides
Abstract : Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) has become an important method to study protein dynamics in solution. Recently, electron-based fragmentation methods (ECD and ETD) have been utilized in HDX-MS/MS experiments as experimental tools to increase the spatial resolution (the ability to obtain deuterium levels of individual residues). An essential prerequisite for this approach is that the level of hydrogen scrambling is negligible. The occurrence of hydrogen scrambling depends critically on the extent of vibrational excitation in the mass spectrometer. In particular, the desolvation process in the electrospray ion source is likely to induce scrambling at standard operating conditions. Consequently, finding experimental conditions that minimize hydrogen scrambling to a negligible level is thus pivotal for the application of electron-based fragmentation in HDX-MS/MS experiments. In the present work, we investigate the occurrence of scrambling in the Apollo I electrospray ion source using ECD of selectively deuterium labeled peptides. The electrospray ion source settings leading to minimal scrambling were identified. Furthermore, an energy dependent loss of deuterium occurring in the ion source was also observed. This loss was critically dependent on the occurrence of scrambling.
https://hal-polytechnique.archives-ouvertes.fr/hal-01048575
Contributor : Denis Roura <>
Submitted on : Friday, July 25, 2014 - 9:26:31 AM
Last modification on : Wednesday, June 9, 2021 - 8:10:01 AM
Contributor : Denis Roura <>
Submitted on : Friday, July 25, 2014 - 9:26:31 AM
Last modification on : Wednesday, June 9, 2021 - 8:10:01 AM