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Spatial-proteomics reveals phospho-signaling dynamics at subcellular resolution

by Frankel, Lisa B. , Lund-Johansen, Fridtjof , Tran, Trung , Østergaard, Ole , Steigerwald, Sophia , Lundby, Alicia , Krogh, Nicolai , Olsen, Jesper V. , Sikorski, Krzysztof , Bekker-Jensen, Simon , Mehta, Adi , Martinez-Val, Ana , Brynjólfsdóttir, Sólveig Hlín , Kjøbsted, Rasmus , Torres-Vega, Estefanía , Kwasniewicz, Ewa , Bekker-Jensen, Dorte B. , Koenig, Claire

in 45 / 631/1647/296 / 631/337/458/1733 / 631/45/475 / 631/80/86/2368 / 82/58 / 96 / 96/63 / Animals / Biological Phenomena / Cell Fractionation / Dynamics / Epidermal growth factor receptors / Fractionation / HeLa Cells / Humanities and Social Sciences / Humans / Hypertonicity / In vivo methods and tests / Localization / Male / Mass Spectrometry / Mass spectroscopy / Mice / Mice, Inbred C57BL / multidisciplinary / Muscle contraction / Muscles / Muscular function / Osmotic Pressure / Phosphorylation / Proteins / Proteome - metabolism / Proteomes / Proteomics / Relocation / Ribosomal proteins / Science / Science (multidisciplinary) / Signal Transduction / Signaling / Subcellular Fractions - metabolism / Workflow

2021

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2021

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2021

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Journal Article

Spatial-proteomics reveals phospho-signaling dynamics at subcellular resolution

Frankel, Lisa B.,

Lund-Johansen, Fridtjof,

Tran, Trung,

Østergaard, Ole,

Steigerwald, Sophia,

Lundby, Alicia,

Krogh, Nicolai,

Olsen, Jesper V.,

Sikorski, Krzysztof,

Bekker-Jensen, Simon,

Mehta, Adi,

Martinez-Val, Ana,

Brynjólfsdóttir, Sólveig Hlín,

Kjøbsted, Rasmus,

Torres-Vega, Estefanía,

Kwasniewicz, Ewa,

Bekker-Jensen, Dorte B.,

Koenig, Claire

2021

Overview

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 . Protein activity regulated by phosphorylation can result in subcellular relocation. Here, the authors present a high throughput spatial phosphoproteomics approach to profile six subcellular compartments, providing insights into EGFR and stress signalling dynamics.

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Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio

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