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Mol Syst Biol DOI:10.15252/msb.202010156

A highly multiplexed quantitative phosphosite assay for biology and preclinical studies.

Publication TypeJournal Article
Year of Publication2021
AuthorsKeshishian, H, E McDonald, R, Mundt, F, Melanson, R, Krug, K, Porter, DA, Wallace, L, Forestier, D, Rabasha, B, Marlow, SE, Jané-Valbuena, J, Todres, E, Specht, H, Robinson, MLea, Beltran, PMJean, Babur, O, Olive, ME, Golji, J, Kuhn, E, Burgess, M, MacMullan, MA, Rejtar, T, Wang, K, Mani, DR, Satpathy, S, Gillette, MA, Sellers, WR, Carr, SA
JournalMol Syst Biol
Date Published2021 Sep

Reliable methods to quantify dynamic signaling changes across diverse pathways are needed to better understand the effects of disease and drug treatment in cells and tissues but are presently lacking. Here, we present SigPath, a targeted mass spectrometry (MS) assay that measures 284 phosphosites in 200 phosphoproteins of biological interest. SigPath probes a broad swath of signaling biology with high throughput and quantitative precision. We applied the assay to investigate changes in phospho-signaling in drug-treated cancer cell lines, breast cancer preclinical models, and human medulloblastoma tumors. In addition to validating previous findings, SigPath detected and quantified a large number of differentially regulated phosphosites newly associated with disease models and human tumors at baseline or with drug perturbation. Our results highlight the potential of SigPath to monitor phosphoproteomic signaling events and to nominate mechanistic hypotheses regarding oncogenesis, response, and resistance to therapy.


Alternate JournalMol Syst Biol
PubMed ID34569154
PubMed Central IDPMC8474009
Grant ListU24 CA210986 / CA / NCI NIH HHS / United States
U24-CA210986 / / HHS | NIH | National Cancer Institute (NCI) /
U24 CA210979 / CA / NCI NIH HHS / United States
U01 CA214125 / CA / NCI NIH HHS / United States
U24CA210979 / / HHS | NIH | National Cancer Institute (NCI) /