Spatial-proteomics reveals phospho-signaling dynamics at subcellular resolution.

Martinez-Val, Ana; Bekker-Jensen, Dorte B; Steigerwald, Sophia; Koenig, Claire; Østergaard, Ole; Mehta, Adi; Tran, Trung; Sikorski, Krzysztof et al. · Nat Commun · 2021

basic_science · Level V

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Abstract

Dynamic change in subcellular localization of signaling proteins is a general concept that eukaryotic cells evolved for eliciting a coordinated response to stimuli. Mass spectrometry-based proteomics in combination with subcellular fractionation can provide comprehensive maps of spatio-temporal regulation of protein networks in cells, but involves laborious workflows that does not cover the phospho-proteome level. Here we present a high-throughput workflow based on sequential cell fractionation to profile the global proteome and phospho-proteome dynamics across six distinct subcellular fractions. We benchmark the workflow by studying spatio-temporal EGFR phospho-signaling dynamics in vitro in HeLa cells and in vivo in mouse tissues. Finally, we investigate the spatio-temporal stress signaling, revealing cellular relocation of ribosomal proteins in response to hypertonicity and muscle contraction. Proteomics data generated in this study can be explored through https://SpatialProteoDynamics.github.io .

Medical subject headings