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Published on: September 20, 2022
Mass Spectrometry-Based Methodologies for Targeted and Untargeted Identification of Protein Covalent Adducts
João Nunes1, Catarina Charneira2, Judit Morello3
1Centro de Química Estrutural, Instituto Superior Técnico, ULisboa, 1049-001 Lisboa, Portugal. joaopcnunes@tecnico.ulisboa.pt.
Abstract:
Protein covalent adducts formed upon exposure to reactive (mainly electrophilic) chemicals may lead to the development of a wide range of deleterious health outcomes. Therefore, the identification of protein covalent adducts constitutes a huge opportunity for a better understanding of events underlying diseases and for the development of biomarkers which may constitute effective tools for disease diagnosis/prognosis, for the application of personalized medicine approaches and for accurately assessing human exposure to chemical toxicants. The currently available mass spectrometry (MS)-based methodologies, are clearly the most suitable for the analysis of protein covalent modifications, providing accuracy, sensitivity, unbiased identification of the modified residue and conjugates along with quantitative information. However, despite the huge technological advances in MS instrumentation and bioinformatics tools, the identification of low abundant protein covalent adducts is still challenging. This review is aimed at summarizing the MS-based methodologies currently used for the identification of protein covalent adducts and the strategies developed to overcome the analytical challenges, involving not only sample pre-treatment procedures but also distinct MS and data analysis approaches.
Insights
Identifying protein covalent adducts is crucial for understanding disease and developing biomarkers. Mass spectrometry (MS) methods are key, but detecting low-abundance adducts remains challenging.
Area of Science:
- Biochemistry
- Toxicology
- Analytical Chemistry
Background:
- Reactive chemicals form protein covalent adducts, potentially causing adverse health effects.
- Identifying these adducts aids disease understanding and biomarker development for diagnosis, personalized medicine, and exposure assessment.
Purpose of the Study:
- To review mass spectrometry (MS)-based methodologies for identifying protein covalent adducts.
- To summarize strategies for overcoming analytical challenges in adduct detection.
Main Methods:
- Utilizes mass spectrometry (MS) for accurate, sensitive, and quantitative analysis of protein modifications.
- Employs advanced bioinformatics tools for data analysis.
Main Results:
- MS-based methods offer high accuracy, sensitivity, and quantitative insights into protein covalent modifications.
- Despite technological advances, identifying low-abundance adducts remains a significant challenge.
Conclusions:
- MS is the most suitable technique for analyzing protein covalent adducts.
- Overcoming challenges requires optimized sample pre-treatment, MS techniques, and data analysis approaches.
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