The B cell-specific major raft protein, Raftlin, is necessary for the integrity of lipid raft and BCR signal transduction

Kazuko Saeki, Yoshiki Miura, Daisuke Aki, Tomohiro Kurosaki, Akihiko Yoshimura

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Recent evidence indicates that membrane microdomains, termed lipid rafts, have a role in B-cell activation as platforms for B-cell antigen receptor (BCR) signal initiation. To gain an insight into the possible functioning of lipid rafts in B cells, we applied liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methodologies to the identification of proteins that co-purified with lipid rafts of Raji cells. Among these raft proteins, we characterized a novel protein termed Raftlin (raft-linking protein). Like the Src family kinase, Raftlin is localized exclusively in lipid rafts by fatty acylation of N-terminal Gly2 and Cys3, and is co-localized with BCR before and after BCR stimulation. Disruption of the Raftlin gene in the DT40 B-cell line resulted in a marked reduction in the quantity of lipid raft components, including Lyn and ganglioside GM1, while overexpression of Raftlin increased the content of raft protein. Moreover, BCR-mediated tyrosine phosphorylation and calcium mobilization were impaired by the lack of Raftlin and actually potentiated by overexpression of Raftlin. These data suggest that Raftlin plays a pivotal role in the formation and/ or maintenance of lipid rafts, therefore regulating BCR-mediated signaling.

Original languageEnglish
Pages (from-to)3015-3026
Number of pages12
JournalEMBO Journal
Volume22
Issue number12
DOIs
Publication statusPublished - 2003 Jun 16
Externally publishedYes

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Signal transduction
B-Cell Antigen Receptors
Signal Transduction
B-Lymphocytes
Cells
Lipids
Proteins
Membrane Microdomains
G(M1) Ganglioside
Acylation
Electrospray ionization
Phosphorylation
src-Family Kinases
Electrospray Ionization Mass Spectrometry
Liquid chromatography
Tandem Mass Spectrometry
Liquid Chromatography
Mass spectrometry
Tyrosine
Chemical activation

Keywords

  • B cell
  • BCR
  • Lipid raft
  • Phosphorylation

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

The B cell-specific major raft protein, Raftlin, is necessary for the integrity of lipid raft and BCR signal transduction. / Saeki, Kazuko; Miura, Yoshiki; Aki, Daisuke; Kurosaki, Tomohiro; Yoshimura, Akihiko.

In: EMBO Journal, Vol. 22, No. 12, 16.06.2003, p. 3015-3026.

Research output: Contribution to journalArticle

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