Dynamic change in subcellular localization of signaling proteins is a general concept that eukaryotic cells evolved for responding and elicit a coordinated response to stimuli. Mass spectrometry (MS)-based proteomics in combination with subcellular fractionation can provide comprehensive maps of spatio-temporal regulation of cells but this is highly challenging involving laborious workflows that does not cover the phosphoproteome level. Here we present a high-throughput workflow based on sequential cell fractionation to profile the global (phospho)proteome dynamics across six distinct subcellular fractions. We benchmarked the workflow by studying spatio-temporal EGFR phospho-signaling dynamics in-vitro in HeLa cells and in-vivo in mouse tissues. Finally, we investigated the spatio-temporal stress signaling , revealing cellular relocation of ribosomal proteins in response to hypertonicity and muscle contraction.

Through this website, you can easily explore the spatio-temporal dynamics of the datasets previously mentioned. Click on "Explore datasets" and select a project to start your queries.

Disclaimer: this resource is for research purpose only.