Top-Down Detection of Oxidative Protein Footprinting by Collision-Induced Dissociation,Electron-Transfer Dissociation, and Electron-Capture Dissociation

0559847 - MBÚ 2023 RIV US eng J - Journal Article
Yassaghi, Ghazaleh - Kukačka, Zdeněk - Fiala, Jan - Kavan, Daniel - Halada, Petr - Volný, Michael - Novák, Petr
Top-Down Detection of Oxidative Protein Footprinting by Collision-Induced Dissociation,Electron-Transfer Dissociation, and Electron-Capture Dissociation.
Analytical Chemistry. Roč. 94, č. 28 (2022), s. 9993-10002. ISSN 0003-2700. E-ISSN 1520-6882
R&D Projects: GA ČR(CZ) GA19-16084S; GA MŠMT(CZ) ED1.1.00/02.0109
EU Projects: European Commission(XE) 731077 - EU_FT-ICR_MS; European Commission(XE) 823839 - EPIC-XS
Research Infrastructure: CIISB II - 90127
Institutional support: RVO:61388971
Keywords : Fast photochemical oxidation of proteins * top-down * collision-induced dissociation * electron-transfer dissociation * electron-capture dissociation * mass spectrometry
OECD category: Analytical chemistry
Impact factor: 7.4, year: 2022
Method of publishing: Limited access
https://pubs.acs.org/doi/10.1021/acs.analchem.1c05476

Fast photochemical oxidation of proteins (FPOP) footprinting is a structural mass spectrometry method that maps proteins by fast and irreversible chemical reactions. The position of oxidative modification reflects solvent accessibility and site reactivity and thus provides information about protein conforma-tion, structural dynamics, and interactions. Bottom-up mass spectrometry is an established standard method to analyze FPOP samples. In the bottom-up approach, all forms of the protein are digested together by a protease of choice, which results in a mixture of peptides from various subpopulations of proteins with varying degrees of photochemical oxidation. Here, we investigate the possibility to analyze a specifically selected population of only singly oxidized proteins. This requires utilization of more specific top-down mass spectrometry approaches. The key element of any top-down experiment is the selection of a suitable method of ion isolation, excitation, and fragmentation. Here, we employ and compare collision-induced dissociation, electron-transfer dissociation, and electron-capture dissociation combined with multi-continuous accumulation of selected ions. A singly oxidized subpopulation of FPOP-labeled ubiquitin was used to optimize the method. The top-down approach in FPOP is limited to smaller proteins, but its usefulness was demonstrated by using it to visualize structural changes induced by co-factor removal from the holo/apo myoglobin system. The top-down data were compared with the literature and with the bottom-up data set obtained on the same samples. The top-down results were found to be in good agreement, which indicates that monitoring a singly oxidized FPOP ion population by the top-down approach is a functional workflow for oxidative protein footprinting.
Permanent Link: https://hdl.handle.net/11104/0334438