Spatial-proteomics reveal in-vivo phospho-signaling dynamics at subcellular resolution

Ana Martinez Del Val, Dorte B Bekker-Jensen, Sophia Steigerwald, Adi Mehta, Trung Tran, Krzysztof Sikorski, Estefania Torres Vega, Ewa Kwasniewicz, Sólveig Hlín Brynjólfsdóttir, Lisa Frankel, Rasmus Kjøbsted, Nicolai Krogh, Alicia Lundby, Simon Bekker-Jensen, Fridtjof Lund-Johansen, Jesper Velgaard Olsen*

*Corresponding author af dette arbejde

Publikation: Working paperPreprintForskning

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 (MS)-based proteomics in combination with subcellular fractionation can provide comprehensive maps of spatio-temporal regulation of 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 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. Proteomics data generated in this study can be explored through https://SpatialProteoDynamics.github.io.
OriginalsprogEngelsk
UdgiverbioRxiv
Sider1-34
Antal sider34
DOI
StatusUdgivet - 2 feb. 2021

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(Preprint)

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