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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/20686

Title: Understanding reaction pathways in top-down ETD by dissecting isotope distributions: A mammoth task
Authors: Lermyte, Frederik
Lacki, Mateusz Krzysztof
Valkenborg, Dirk
Baggerman, Geert
Gambin, Anna
Sobott, Frank
Issue Date: 2015
Publisher: ELSEVIER SCIENCE BV
Citation: INTERNATIONAL JOURNAL OF MASS SPECTROMETRY, 390, p. 146-154
Abstract: Electron transfer dissociation (ETD) is becoming increasingly more important in mass spectrometrybased structural proteomics due to its ability to cleave the protein backbone without annihilating the higher-order structure. Concomitant with ETD, non-dissociative charge reduction occurs primarily by proton transfer from the protein precursor to the ETD reagent, and by non-dissociative electron transfer from the reagent to the protein. While it has been known for some time that the preference for either proton or electron transfer is largely a function of the properties of the reagent, the extent of dissociative versus non-dissociative electron transfer is also dependent on instrument choice, precursor charge state and conformation, as well as ion activation before, during, and after cation/anion interaction. In this work, we use a novel software named MASSTODON to quantify the different reaction pathways in top-down spectra of ubiquitin, acquired on two different instruments: a Synapt G2 (Waters) and an LTQ Orbitrap Velos (Thermo). We compare the behavior of different precursor charge states, representing more folded or more extended conformations, and also evaluate the effect of increasing supplemental activation (on the Synapt) and reaction time (on the Orbitrap). In agreement with previous work, increasing supplemental activation promotes fragment release and hydrogen radical migration from (N-terminal) c fragments to (C-terminal) z fragments. A longer reaction time leads to increasingly more extensive charge reduction and a decrease in the relative occurrence of non-dissociative electron transfer compared to proton transfer, possibly indicating some degree of time- andtharge-state dependent loss of electrostatic interactions on a timescale of a few tens of milliseconds. (C) 2015 Elsevier B.V. All rights reserved.
Notes: [Lermyte, Frederik; Sobott, Frank] Univ Antwerp, Dept Chem, Biomol & Analyt Mass Spectrometry Grp, B-2020 Antwerp, Belgium. [Lermyte, Frederik; Valkenborg, Dirk; Baggerman, Geert; Sobott, Frank] Univ Antwerp, UA VITO Ctr Prote, B-2020 Antwerp, Belgium. [Lacki, Mateusz Krzysztof; Gambin, Anna] Univ Warsaw, Inst Informat, PL-00325 Warsaw, Poland. [Valkenborg, Dirk] Hasselt Univ, Interuniv Inst Biostat & Stat Bioinformat, Hasselt, Belgium. [Valkenborg, Dirk; Baggerman, Geert] Flemish Inst Technol Res VITO, Appl Bio & Mol Syst, Antwerp, Belgium.
URI: http://hdl.handle.net/1942/20686
DOI: 10.1016/j.ijms.2015.08.008
ISI #: 000366079600019
ISSN: 1387-3806
Category: A1
Type: Journal Contribution
Validation: ecoom, 2017
Appears in Collections: Research publications

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