Temporal proteomics profiling of lipid rafts in CCR6-activated T cells reveals the integration of actin cytoskeleton dynamics

Shu Ling Lin, Chih Wei Chien, Chia Li Han, Eric S W Chen, Shao Hsuan Kao, Yu Ju Chen, Fang Liao

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Chemokines orchestrate leukocyte migration toward sites of inflammation and infection and target leukocytes via chemokine receptors such as CCR6, a subfamily of the seven-transmembrane G-proteincoupled receptors. Lipid rafts are cholesterol and sphingolipid-enriched liquid-ordered membrane microdomains thought to serve as scaffolding platforms for signal transduction. To globally understand the dynamic changes of proteins within lipid rafts upon CCR6 activation in T cells, we quantitatively analyzed the time-dependent changes of lipid raft proteome using our recently reported membrane proteomics strategy combining gel-assisted digestion, iTRAQ labeling and LC-MS/MS. To our knowledge, the error-free identification of 852 proteins represents the first data set of the raft proteome in T cells upon chemokine receptor activation, including 354 previously annotated raft proteins and 85 dynamically recruited proteins that are potential raft-associated proteins. The temporal profiles revealed that many proteins involved in the actin cytoskeleton rearrangement are actively recruited into lipid rafts upon CCR6 activation. We further confirmed the proteomics results by Western blotting and used small interfering RNA-mediated knockdown to evaluate their roles upon CCR6 activation. In sum, we employed quantitative proteomic strategy to analyze raft proteome and identified manymolecules actively involved in the control of actin assembly and disassembly regulating CCR6 activation-induced cell migration.

Original languageEnglish
Pages (from-to)283-297
Number of pages15
JournalJournal of Proteome Research
Volume9
Issue number1
DOIs
Publication statusPublished - Jan 4 2010
Externally publishedYes

Keywords

  • CCR6
  • G-protein-coupled receptors
  • Gel-assisted digestion
  • iTRAQ
  • Lipid raft proteome
  • Mass spectrometry
  • T cells

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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