TY - JOUR
T1 - Spatial-proteomics reveals phospho-signaling dynamics at subcellular resolution
AU - Martinez-Val, Ana
AU - Bekker-Jensen, Dorte B.
AU - Steigerwald, Sophia
AU - Koenig, Claire
AU - Østergaard, Ole
AU - Mehta, Adi
AU - Tran, Trung
AU - Sikorski, Krzysztof
AU - Torres-Vega, Estefanía
AU - Kwasniewicz, Ewa
AU - Brynjólfsdóttir, Sólveig Hlín
AU - Frankel, Lisa B.
AU - Kjøbsted, Rasmus
AU - Krogh, Nicolai
AU - Lundby, Alicia
AU - Bekker-Jensen, Simon
AU - Lund-Johansen, Fridtjof
AU - Olsen, Jesper V.
N1 - © 2021. The Author(s).
PY - 2021
Y1 - 2021
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-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 .
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-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 .
U2 - 10.1038/s41467-021-27398-y
DO - 10.1038/s41467-021-27398-y
M3 - Journal article
C2 - 34876567
VL - 12
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 7113
ER -