{Reference Type}: Journal Article
{Title}: Multifaceted Proteome Analysis at Solubility, Redox, and Expression Dimensions for Target Identification.
{Author}: Saei AA;Lundin A;Lyu H;Gharibi H;Luo H;Teppo J;Zhang X;Gaetani M;Végvári Á;Holmdahl R;Gygi SP;Zubarev RA;
{Journal}: Adv Sci (Weinh)
{Volume}: 0
{Issue}: 0
{Year}: 2024 Aug 9
{Factor}: 17.521
{DOI}: 10.1002/advs.202401502
{Abstract}: Multifaceted interrogation of the proteome deepens the system-wide understanding of biological systems; however, mapping the redox changes in the proteome has so far been significantly more challenging than expression and solubility/stability analyses. Here, the first high-throughput redox proteomics approach integrated with expression analysis (REX) is devised and combined with the Proteome Integral Solubility Alteration (PISA) assay. The whole PISA-REX experiment with up to four biological replicates can be multiplexed into a single tandem mass tag TMTpro set. For benchmarking this compact tool, HCT116 cells treated with auranofin are analyzed, showing great improvement compared with previous studies. PISA-REX is then applied to study proteome remodeling upon stimulation of human monocytes by interferon α (IFN-α). Applying this tool to study the proteome changes in plasmacytoid dendritic cells (pDCs) isolated from wild-type versus Ncf1-mutant mice treated with interferon α, shows that NCF1 deficiency enhances the STAT1 pathway and modulates the expression, solubility, and redox state of interferon-induced proteins. Providing comprehensive multifaceted information on the proteome, the compact PISA-REX has the potential to become an industry standard in proteomics and to open new windows into the biology of health and disease.