A Large-Scale Quantitative Proteomic Approach To Identifying Sulfur Mustard-Induced Protein Phosphorylation Cascades

Abstract

Sulfur mustard [SM, bis-(2-chloroethyl) sulfide] is a potent alkylating agent and chemical weapon. While there are no effective treatments for SM-induced injury, current research focuses on understanding the molecular changes upon SM exposure. Indeed, efforts that seek a more comprehensive analysis of proteins and post-translational modifications are critical for understanding SM-induced toxicity on a more global scale. Furthermore, these studies can uncover proteins previously uncharacterized in SM-exposed cells, which in turn leads to potential targets for therapeutic intervention. Here, we apply a quantitative proteomic approach termed stable isotope-labeling with amino acids in cell culture combined with immobilized metal affinity chromatography to study the large-scale protein phosphorylation changes resulting from SM exposure in a human keratinocyte cell culture model. This resulted in the characterization of over 2300 nonredundant phosphorylation sites, many of which exhibit altered levels in response to SM. Our results point toward several proteins previously implicated in SM-induced toxicity as well as many additional proteins previously uncharacterized. Further de novo analysis of these phosphoproteins using interaction mapping software revealed both known and novel pathways that may serve as future therapeutic targets of SM exposure.

A review on symptoms, treatments protocols, and proteomic profile in sulfur mustard‐exposed victims

“…In 2010, Everley et al showed that guanine nucleotide exchange factors (GEFs) and GTPase‐activating proteins (GAPs) were down‐regulated by up to two folds in an in vitro study on human keratinocytes model after SM‐exposure. However phosphorylated proteins showed up to threefold increases in the SM‐exposed cells …”

Transient Receptor Potential Channel A1 (TRPA1) Regulates Sulfur Mustard-Induced Expression of Heat Shock 70 kDa Protein 6 (HSPA6) In Vitro

“…Both cell types were used in several studies before and were found very well suited for the investigation of TRPA1-related effects [ 23 , 40 , 41 ]. Several genes and proteins have been reported to be specifically up-regulated in mouse skin and in human keratinocytes after exposure to SM [ 42 , 43 ]. Thus, we focused on proteome changes after SM exposure with special focus on the involvement of TRPA1.…”

Sulfur mustard induced nuclear translocation of glyceraldehyde-3-phosphate-dehydrogenase (GAPDH)

“…In addition, comprehensive analyses of proteins and posttranslational protein modifications have contributed to unveil the complex pathophysiology of SM and have given new impulses for the development of new potential therapeutic measurements [14][15][16].…”