Thiol redox proteomics seen with fluorescent eyes: The detection of cysteine oxidative modifications by fluorescence derivatization and 2-DE

• Published in: Journal of proteomics Vol. 75; no. 2; pp. 329 - 338
• Main Authors: Izquierdo-Álvarez, Alicia, Martínez-Ruiz, Antonio
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 21.12.2011; Elsevier
• Subjects: 2-DE; Animals; Biological and medical sciences; cell communication; cysteine; Cysteine - chemistry; Cysteine oxidation; derivatization; Diverse techniques; Electrophoresis, Gel, Two-Dimensional - methods; eyes; fluorescence; Fluorescent Dyes - chemistry; Fundamental and applied biological sciences. Psychology; Humans; Molecular and cellular biology; Oxidation-Reduction; Post-translational modifications; Protein Processing, Post-Translational; Proteins - chemistry; proteome; proteomics; Proteomics - methods; Redox DIGE; Redox fluorescence switch (RFS); spectrometers; Sulfhydryl Compounds - chemistry; Sulfhydryl Compounds - metabolism; Thiol redox proteomics; thiols; two-dimensional gel electrophoresis; Post-translational modifications; Cysteine oxidation; Thiol redox proteomics; 2-DE; Redox fluorescence switch (RFS); Redox DIGE; Cysteine; Thiol; Fluorescence; Posttranslational modification; Eye; Proteomics; Oxidation; Detection
• ISSN: 1874-3919; 1876-7737
• DOI: 10.1016/j.jprot.2011.09.013

There is increasing evidence that several reversible oxidative post-translational modifications of protein cysteines participate in cell signalling. Specific proteomic techniques are required to identify these modifications and to study their regulation in different cell processes, that are collectively known as thiol redox proteomics. Recently, fluorescence derivatization methods have been developed that enable these post-translational modifications to be studied using proteomic workflows based on two-dimensional electrophoresis, which is a relatively accessible and affordable technique. As well as enabling a large number of samples to be processed, two-dimensional electrophoresis has the advantage that it does not rely on the intensive use of mass spectrometers. This methodology allows to “visualise” redox changes in a broad context and, although identification of the modified residues is not so straightforward, complementary derivatization can overcome this drawback. Here we review the different derivatization strategies that have been employed in these studies, comparing their advantages and potential limitations. We also review the applications and results obtained, with particular emphasis on those involving (patho)physiological stimuli, thereby showing the potential of these techniques to study the thiol redox proteome. [Display omitted] ► Reversible protein cysteine oxidation is now a key issue in cell signalling. ► Fluorescence derivatization coupled to 2DE allows to study these modifications. ► We critically review methodological differences in the techniques used. ► They have allowed to study protein Cys oxidation in some physiological scenarios.