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Series GSE41490 Query DataSets for GSE41490
Status Public on Apr 11, 2013
Title Direct proteomic quantification of the secretome of activated immune cells
Organism Mus musculus
Experiment type Expression profiling by array
Summary Protein secretion can mediate communication of distant cells in a multicellular organism and can control a broad range of physiological functions. Regulation of this process is crucial, especially for cells of the immune system that sense pathogens through specific receptors and release proteins that initiate and orchestrate the immune response. This secretome is today analyzed using antibody-based assays, but mass spectrometry could provide an unbiased and comprehensive alternative. Here we developed a quantitative, high resolution mass spectrometric workflow to detect and quantify proteins that are released from macrophages upon Toll-like receptor 4 (TLR4) activation by the microbial component lipopolysaccharide (LPS)5. We quantified the time-resolved release of 775 proteins, including 52 annotated cytokines, from only 150.000 primary cells per condition. The dynamic range of the secretory induction was at least 10,000 fold and we achieved sensitivity in the low picogram range. Comparison to the transcriptome revealed high correlation for some protein classes but also anti-correlation resulting from the transcriptionally decoupled release of lysosomal proteins. Using gene knockout (KO) mouse strains as genetic models with intracellular signaling adaptor deficiencies, we defined distinct secretory profiles depending on which arms of the signaling pathways are missing. We show that MyD88 dominates over TRIF in the number and fold-induction of secreted proteins6,7. Interestingly, the sum of the two signaling arms does not equal the wild-type response; instead, protein secretion is tuned by synergistic and predominantly redundant mechanisms. The kinetic data reveals that the release of anti-inflammatory proteins dampens the initial pro-inflammatory response. Our study provides a paradigm for the sensitive and time-resolved identification of receptor activation induced cellular secretomes, including proteins with unexpected extracellular presence, and allows the characterization of the underlying signal transduction pathways.
 
Overall design Microarray experiments were performed as dual-color hybridizations.
 
Contributor(s) Meissner F, Scheltema RA, Mann M, Mollenkopf H
Citation(s) 23620052
Submission date Oct 11, 2012
Last update date Jan 19, 2018
Contact name Hans-Joachim Mollenkopf
E-mail(s) mollenkopf@mpiib-berlin.mpg.de
Phone +49 30 28460 482
Organization name Max-Planck-Institute for Infection Biology
Lab Microarray/Genomics Core Facility
Street address Charitéplatz 1
City Berlin
ZIP/Postal code 10117
Country Germany
 
Platforms (1)
GPL10333 Agilent-026655 Whole Mouse Genome Microarray 4x44K v2 (Feature Number version)
Samples (20)
GSM1018150 WT 1 (1h) - vs. WT 1 (1h) LPS
GSM1018151 WT 2 (2h) - vs. WT 2 (2h) LPS
GSM1018152 WT 3 (4h) - vs. WT 3 (4h) LPS
Relations
BioProject PRJNA177213

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE41490_RAW.tar 473.9 Mb (http)(custom) TAR (of TXT, XML)
Processed data included within Sample table
Processed data provided as supplementary file

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