Quantitation of phosphorylation levels in the TLR signaling pathway in mouse macrophages using iTRAQ

Tuesday, October 25, 2011 — Poster Session II
Noon – 2:00 p.m.	Natcher Conference Center	NIAID	PROTEOM-6

Authors

• V Sjoelund
• A Nita-Lazar

Abstract

The Toll-like receptors (TLRs) are a family of pathogen recognition receptors that alert the host to the presence of pathogens. Cytokines are among the most important genes to be regulated by TLR signaling, and given their role in the orchestration of the inflammatory response, mechanisms of modulating their production has garnered a lot of interest, in particular in the area of the development of therapies for the treatment of chronic inflammatory diseases. Using a phosphoproteomics approach, we searched for proteins whose phosphorylation levels change post activation of the TLR pathway. Mouse macrophages were activated with lipopolysaccharide (LPS) and the levels of phosphorylation at different time points post activation were quantified using an iTRAQ 8-plex reagent on an Orbitrap Velos. We have identified 47 proteins that have significant changes in phosphorylation levels at different time points post activation. Most of these proteins were cytoskeletal proteins or transcription factors. Considering the major morphology changes that the macrophages need to undergo for function (phagocytosis for example) and that the major role of the TLR pathway is to turn on genes, those results confirm the role of phosphorylation in the rapid transmission of the LPS signal.

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