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Enhancing Biomolecule Analysis and 2DMS Experiments by Implementation of (Activated Ion) 193 nm UVPD on a FT-ICR Mass Spectrometer
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Theisen, Alina, Wootton, Christopher, Haris, Anisha, Morgan, Tomos E., Lam, Yuko P. Y., Barrow, Mark P. and O’Connor, Peter B. (2022) Enhancing Biomolecule Analysis and 2DMS Experiments by Implementation of (Activated Ion) 193 nm UVPD on a FT-ICR Mass Spectrometer. Analytical Chemistry, 94 (45). pp. 15631-15638. doi:10.1021/acs.analchem.2c02354 ISSN 1520-6882.
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Official URL: https://doi.org/10.1021/acs.analchem.2c02354
Abstract
Ultraviolet photodissociation is a fast, photon-mediated fragmentation method that yields high sequence coverage and informative cleavages of biomolecules. In this work, 193 nm UVPD was coupled with a 12 Tesla FT-ICR mass spectrometer and 10.6 μm infrared multi-photon dissociation to provide gentle slow-heating of UV-irradiated ions. No internal instrument hardware modifications were required. Adjusting the timing of laser pulses to the ion motion within the ICR cell provided consistent fragmentation yield shot-to-shot and may also be used to monitor ion positions within the ICR cell. Single-pulse UVPD of the native-like 5+ charge state of ubiquitin resulted in 86.6% cleavage coverage. Additionally, IR activation post UVPD doubled the overall fragmentation yield and boosted the intensity of UVPD-specific x-type fragments up to 4-fold. This increased yield effect was also observed for the 6+ charge state of ubiquitin, albeit less pronounced. This indicates that gentle slow-heating serves to sever tethered fragments originating from non-covalently linked compact structures and makes activation post UVPD an attractive option to boost fragmentation efficiency for top-down studies. Lastly, UVPD was implemented and optimized as a fragmentation method for 2DMS, a data-independent acquisition method. UVPD-2DMS was demonstrated to be a viable method using BSA digest peptides as a model system.
Item Type: | Journal Article | |||||||||||||||
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Subjects: | Q Science > QD Chemistry T Technology > TP Chemical technology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | |||||||||||||||
SWORD Depositor: | Library Publications Router | |||||||||||||||
Library of Congress Subject Headings (LCSH): | Photodissociation, Biomolecules -- Separation, Mass spectrometry | |||||||||||||||
Journal or Publication Title: | Analytical Chemistry | |||||||||||||||
Publisher: | American Chemical Society | |||||||||||||||
ISSN: | 1520-6882 | |||||||||||||||
Official Date: | 1 November 2022 | |||||||||||||||
Dates: |
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Volume: | 94 | |||||||||||||||
Number: | 45 | |||||||||||||||
Page Range: | pp. 15631-15638 | |||||||||||||||
DOI: | 10.1021/acs.analchem.2c02354 | |||||||||||||||
Status: | Peer Reviewed | |||||||||||||||
Publication Status: | Published | |||||||||||||||
Access rights to Published version: | Open Access (Creative Commons) | |||||||||||||||
Date of first compliant deposit: | 5 December 2022 | |||||||||||||||
Date of first compliant Open Access: | 5 December 2022 | |||||||||||||||
RIOXX Funder/Project Grant: |
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