Luise, Arianna and De Cecco, Elena and Ponzini, Erika and Sollazzo, Martina and Mauri, PierLuigi and Sobott, Frank and Legname, Giuseppe and Grandori, Rita and Santambrogio, Carlo (2021) Profiling Dopamine-Induced Oxidized Proteoforms of β-synuclein by Top-Down Mass Spectrometry. Antioxidants, 10 (6). p. 893. ISSN 2076-3921
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Abstract
The formation of multiple proteoforms by post-translational modifications (PTMs) enables a single protein to acquire distinct functional roles in its biological context. Oxidation of methionine residues (Met) is a common PTM, involved in physiological (e.g., signaling) and pathological (e.g., oxidative stress) states. This PTM typically maps at multiple protein sites, generating a heterogeneous population of proteoforms with specific biophysical and biochemical properties. The identification and quantitation of the variety of oxidized proteoforms originated under a given condition is required to assess the exact molecular nature of the species responsible for the process under investigation. In this work, the binding and oxidation of human β-synuclein (BS) by dopamine (DA) has been explored. Native mass spectrometry (MS) has been employed to analyze the interaction of BS with DA. In a second step, top-down fragmentation of the intact protein from denaturing conditions has been performed to identify and quantify the distinct proteoforms generated by DA-induced oxidation. The analysis of isobaric proteoforms is approached by a combination of electron-transfer dissociation (ETD) at each extent of modification, quantitation of methionine-containing fragments and combinatorial analysis of the fragmentation products by multiple linear regression. This procedure represents a promising approach to systematic assessment of proteoforms variety and their relative abundance. The method can be adapted, in principle, to any protein containing any number of methionine residues, allowing for a full structural characterization of the protein oxidation states.
Item Type: | Article |
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Subjects: | STM One > Agricultural and Food Science |
Depositing User: | Unnamed user with email support@stmone.org |
Date Deposited: | 17 Jul 2023 04:37 |
Last Modified: | 11 May 2024 09:56 |
URI: | http://publications.openuniversitystm.com/id/eprint/1663 |