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Title: | Characterization of top-down ETD in a travelling-wave ion guide | Authors: | Lermyte, Frederik Verschueren, Tim Brown, Jeffery M. Williams, Jonathan P. VALKENBORG, Dirk Sobott, Frank |
Issue Date: | 2015 | Publisher: | ACADEMIC PRESS INC ELSEVIER SCIENCE | Source: | METHODS, 89, p. 22-29 | Abstract: | Top-down sequencing methods are becoming increasingly relevant for protein characterization, in particular electron capture (ECD) and electron transfer dissociation (ETD) which allow for extensive backbone cleavage with minimal side reactions. The ability to obtain sequence-specific fragments while maintaining aspects of the higher-order structure, as well as the position of deuterium labels in HID exchange, has attracted interest from scientists in the field of structural proteomics. Recently, ETD has also been combined with ion mobility on commercially available quadrupole/time-of-flight instruments, and this implementation paves the way to novel structural studies and investigation of the ETD process itself. In the current work, we investigate the use of ETD for fragmentation of standard peptides and proteins and provide a detailed description of the effect of the parameters controlling the time and efficiency of the reaction. We also highlight how the combination with ion mobility separation after electron transfer provides extended analytical benefits, such as assignment of fragments to a specific charge-reduced state of the precursor. (C) 2015 Elsevier Inc. All rights reserved. | Notes: | [Lermyte, Frederik; Verschueren, Tim; Sobott, Frank] Univ Antwerp, Dept Chem, Biomol & Analyt Mass Spectrometry Grp, B-2020 Antwerp, Belgium. [Lermyte, Frederik; Valkenborg, Dirk; Sobott, Frank] Univ Antwerp, UA VITO Ctr Prote, B-2020 Antwerp, Belgium. [Brown, Jeffery M.; Williams, Jonathan P.] Waters Corp, Wilmslow SK9 4AX, Cheshire, England. [Valkenborg, Dirk] Hasselt Univ, Interuniv Inst Biostat & Stat Bioinformat, Hasselt, Belgium. [Valkenborg, Dirk] Flemish Inst Technol Res VITO, Appl Bio & Mol Syst, Mol, Belgium. | Keywords: | Top-down mass spectrometry; Intact protein mass spectrometry; Top-down sequencing; Electron transfer dissociation; Ion mobility;top-down mass spectrometry; intact protein mass spectrometry; top-down sequencing; electron transfer dissociation; ion mobility | Document URI: | http://hdl.handle.net/1942/20601 | ISSN: | 1046-2023 | e-ISSN: | 1095-9130 | DOI: | 10.1016/j.ymeth.2015.05.019 | ISI #: | 000365062800004 | Rights: | © 2015 Elsevier Inc. All rights reserved. | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2016 |
Appears in Collections: | Research publications |
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