TY - UNPB
T1 - Spatial-proteomics reveal in-vivo phospho-signaling dynamics at subcellular resolution
AU - Martinez Del Val, Ana
AU - Bekker-Jensen, Dorte B
AU - Steigerwald, Sophia
AU - Mehta, Adi
AU - Tran, Trung
AU - Sikorski, Krzysztof
AU - Torres Vega, Estefania
AU - Kwasniewicz, Ewa
AU - Brynjólfsdóttir, Sólveig Hlín
AU - Frankel, Lisa
AU - Kjøbsted, Rasmus
AU - Krogh, Nicolai
AU - Lundby, Alicia
AU - Bekker-Jensen, Simon
AU - Lund-Johansen, Fridtjof
AU - Olsen, Jesper Velgaard
N1 - (Preprint)
PY - 2021/2/2
Y1 - 2021/2/2
N2 - 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.
AB - 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.
U2 - 10.1101/2021.02.02.425898
DO - 10.1101/2021.02.02.425898
M3 - Preprint
SP - 1
EP - 34
BT - Spatial-proteomics reveal in-vivo phospho-signaling dynamics at subcellular resolution
PB - bioRxiv
ER -