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 - Článek v odborném periodiku
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
Grant CEP: GA ČR(CZ) GA19-16084S; GA MŠMT(CZ) ED1.1.00/02.0109
GRANT EU: European Commission(XE) 731077 - EU_FT-ICR_MS; European Commission(XE) 823839 - EPIC-XS
Výzkumná infrastruktura: CIISB II - 90127
Institucionální podpora: RVO:61388971
Klíčová slova: Fast photochemical oxidation of proteins * top-down * collision-induced dissociation * electron-transfer dissociation * electron-capture dissociation * mass spectrometry
Obor OECD: Analytical chemistry
Impakt faktor: 7.4, rok: 2022
Způsob publikování: Omezený přístup
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.
Trvalý link: https://hdl.handle.net/11104/0334438