Proteome analysis of lipid rafts in Jurkat cells characterizes a raft subset that is involved in NF-κB activation

Xiaolin Tu, Aaron Huang, David Bae, Ndaisha Slaughter, Julian Whitelegge, Timothy Crother, Perry E. Bickel, Andre Nel

Research output: Contribution to journalArticle

31 Scopus citations

Abstract

Lipid rafts are detergent-insoluble membrane domains that play a key role in signal transduction by the T-cell antigen receptor. Proteome analysis revealed the presence of amidosulfobetaine-soluble signal transducing, integral membrane, cytoskeletal, heat shock, and GTP-binding proteins in rafts prepared from Jurkat cells. Several of these proteins were recruited to rafts by CD3/CD28 costimulation. Of particular interest is the inducible association of activated IκB kinase complexes with raft vesicles that could be captured with anti-flotillin-1 antibodies. Following amidosulfobetaine solubilization, flotillin-1 and IKKβ underwent reciprocal co-immunoprecipitation. Treatment of Jurkat cells with methyl-β-cyclodextrin disrupted the assembly and activation of this raft complex and also interfered in CD3/ CD28-induced activation of a NF-κB response element in the IL-2 promoter.

Original languageEnglish (US)
Pages (from-to)445-454
Number of pages10
JournalJournal of Proteome Research
Volume3
Issue number3
DOIs
StatePublished - May 1 2004

Keywords

  • Lipid rafts
  • NF-κB signaling
  • Proteomics
  • T-cell antigen receptor

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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