Glutaredoxin 1 protects lens epithelial cells from epithelial-mesenchymal transition by preventing casein kinase 1α S-glutathionylation during posterior capsular opacification

Li, Chenshuang; Chen, Xi; Zhang, Siqi; Chen, Liang; Ma, Xiao‐Pan; Zhang, Ruixue; Zhang, Pengcheng

doi:10.1016/j.redox.2023.102676

Cited by 4 publications

(1 citation statement)

References 56 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mass spectrometry (MS) and proteomics have been instrumental in the identification, validation, and discovery of SSG proteins and cysteines (Kukulage et al., 2022). In the field of SSG, experimentalists seek to discover new SSG proteins via proteomics to understand a phenotype of interest or analyze pathways through system biology investigations (Guo et al., 2021; Li et al., 2023). In addition, there is interest in validating the SSG of a protein of interest (POI) in a given biological model (Adachi et al., 2004; Alegre‐Cebollada et al., 2014).…”

Section: Introductionmentioning

confidence: 99%

Clickable Glutathione‐Based Identification of Cysteine Glutathionylation

Matarage Don,

Kukulage,

Ahn

2023

Current Protocols

View full text Add to dashboard Cite

Clickable glutathione is a glutathione‐derived chemical probe designed to identify and analyze protein S‐glutathionylation, a major cysteine oxidation in redox signaling. An engineered glutathione synthetase mutant (GS M4) is used to synthesize clickable glutathione in cells or in vitro, which affords utility via click chemistry to detect, identify, and quantify glutathionylation on individual or global proteins in biochemical and mass spectrometric analyses. The clickable glutathione approach is valuable for the unequivocal identification of glutathionylated cysteines, among many reversible cysteine oxoforms, via the direct enrichment and detection of glutathionylated proteins or peptides. Clickable glutathione, in combination with GS M4, has demonstrated utility in the mass‐spectrometry‐based discovery and profiling of new proteins and cysteines for glutathionylation in cell lines in response to physiologic and oxidative stress. The approach is versatile and applicable to validating the glutathionylation of proteins and cysteines in other biochemical analysis beside mass spectrometry. Here, we describe the applications of clickable glutathione and provide detailed protocols for the identification, profiling, and detection of glutathionylated proteins and cysteines. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Identification of glutathionylated cysteine in individual proteins in vitroBasic Protocol 2: Proteomic identification and quantification of glutathionylationBasic Protocol 3: Biochemical validation of glutathionylation in cells

show abstract