Multifaceted Proteome Analysis at Solubility, Redox, and Expression Dimensions for Target Identification.

Autor: Saei AA; Department of Cell Biology, Harvard Medical School, Boston, MA, 02115, USA.; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden.; Biozentrum, University of Basel, Basel, 4056, Switzerland.; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, 17165, Sweden., Lundin A; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden., Lyu H; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden., Gharibi H; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden., Luo H; Division of Immunology, Medical Inflammation Research Group, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, SE-17 177, Sweden., Teppo J; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden.; Drug Research Program, Faculty of Pharmacy, University of Helsinki, Helsinki, FI-00014, Finland., Zhang X; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden., Gaetani M; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden.; SciLifeLab, Stockholm, SE-17 177, Sweden., Végvári Á; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden., Holmdahl R; Division of Immunology, Medical Inflammation Research Group, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, SE-17 177, Sweden., Gygi SP; Department of Cell Biology, Harvard Medical School, Boston, MA, 02115, USA., Zubarev RA; Division of Chemistry I, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, SE-17 177, Sweden.; SciLifeLab, Stockholm, SE-17 177, Sweden.
Jazyk: angličtina
Zdroj: Advanced science (Weinheim, Baden-Wurttemberg, Germany) [Adv Sci (Weinh)] 2024 Oct; Vol. 11 (38), pp. e2401502. Date of Electronic Publication: 2024 Aug 09.
DOI: 10.1002/advs.202401502
Abstrakt: 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.
(© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)
Databáze: MEDLINE
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