Speaker: Prof. Jesper Velgaard Olsen - Vice Director, University of Copenhagen, Faculty of Health and Medical Sciences / Novo Nordisk Foundation Center for Protein Research, Proteomics Program, Denmark
Host: IRB Proteomics Data Club (PDC)
Organizer: IRB Research Nodes
Date: Wednesday 10 March 2021, 12.00h
Dynamic changes in subcellular localization of signaling proteins are a general concept that eukaryotic cells make use of to transmit information about extracellular cues to the nucleus and elicit a coordinated response to the stimuli. We developed a streamlined and high-throughput spatial proteomics approach for analyzing subcellular phosphoproteome dynamics in a temporal manner. The method is based on sequential cell fractionation using different biochemical buffers to profile six distinct subcellular fractions with high reproducibility. Fast chromatographic separation using the Evosep One coupled to directDIA measuremens on an Orbitrap Exploris 480 mass spectrometer allows acquisition of complete data for both subcellular proteome and phosphoproteome in just 4 hours of MS time. Spatio-temporal analysis of the osmotic shock response and EGF signaling in cancer cells and in-vivo in mouse tissues reveals new interesting insights into subcellular phosphoprotein dynamics.