Redox proteomics in melanoma cells: An optimized protocol.

Autor: Cunha ES; Departamento de Bioquímica e Biologia Molecular, Programa de Pós-graduação Em Ciências (Bioquímica), UFPR, Brazil., Mazepa E; Departamento de Bioquímica e Biologia Molecular, Programa de Pós-graduação Em Ciências (Bioquímica), UFPR, Brazil., Batista M; Laboratório de Ciências e Tecnologias Aplicadas Em Saúde (LaCTAS), Instituto Carlos Chagas - FIOCRUZ/PR, Brazil., Marchini FK; Laboratório de Ciências e Tecnologias Aplicadas Em Saúde (LaCTAS), Instituto Carlos Chagas - FIOCRUZ/PR, Brazil., Martinez GR; Departamento de Bioquímica e Biologia Molecular, Programa de Pós-graduação Em Ciências (Bioquímica), UFPR, Brazil. Electronic address: grmartinez@ufpr.br.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2024 Aug; Vol. 691, pp. 115543. Date of Electronic Publication: 2024 Apr 16.
DOI: 10.1016/j.ab.2024.115543
Abstrakt: Cancer development and progression are intimately related with post-translational protein modifications, e.g., highly reactive thiol moiety of cysteines enables structural rearrangements resulting in redox biological switches. In this context, redox proteomics techniques, such as 2D redox DIGE, biotin switch assay and OxIcat are fundamental tools to identify and quantify redox-sensitive proteins and to understand redox mechanisms behind thiol modifications. Given the great variability in redox proteomics protocols, problems including decreased resolution of peptides and low protein amounts even after enrichment steps may occur. Considering the biological importance of thiol's oxidation in melanoma, we adapted the biotin-switch assay technique for melanoma cells in order to overcome the limitations and improve coverage of detected proteins.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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