Stimulation of the high-affinity IgE receptor results in the tyrosine phosphorylation of a 60 kD protein which is associated with the protein- tyrosine kinase, Csk

Thorunn Rafnar, R. Stokes Peebles, Mary E. Brummet, Branimir Čatipović, Farhad Imani, Donald W. MacGlashan, David G. Marsh

Research output: Contribution to journalArticlepeer-review

Abstract

The protein tyrosine kinase Csk downregulates the activity of the Src family of kinases and has a negative effect on signal transduction through several Src kinase-associated receptors. Because the Sin-family kinase Lyn plays a pivotal role in FcεRI-mediated cellular activation, we examined whether Csk is involved in FcεRI signaling events. Using anti-Csk antibodies and recombinant fusion proteins we detected a single tyrosine-phosphorylated protein of 60 kD (herein referred to as 'p60') that associates with the SH2 domain of Csk after stimulation of the FcεRI. p.60 phosphorylation reached a maximum within one minute and remained constant while the receptors were aggregated; disaggregation of the receptors resulted in rapid dephosphorylation of p60. The phosphorylation of p60 was only detected after activation by IgE and antigen and not by stimulation with PMA and/or ionomycin. Phosphorylated p60 was associated entirely with the membrane fraction of the cells. A considerable fraction of Csk was associated with the membrane in both unstimulated and stimulated cells, this fraction did not change upon activation. p60 coprecipitated with Csk from both unstimulated and FcεRI stimulated cells and was phosphorylated by the immunocomplex. Total kinase activity of Csk immunoprecipitates increased upon FcεRI stimulation. p60 did not react with antibodies to a number of known signaling molecules, including the recently cloned, GAP-associated protein, p62(dok). Our data demonstrate that Csk associates with a membrane-anchored protein complex that is directly involved in FcεRI signal transduction.

Original languageEnglish (US)
Pages (from-to)249-257
Number of pages9
JournalMolecular Immunology
Volume35
Issue number4
DOIs
StatePublished - Mar 1 1998

Keywords

  • Fc receptors
  • Mast cells/basophils
  • Protein kinases/phosphatases
  • Signal transduction

ASJC Scopus subject areas

  • Immunology
  • Molecular Biology

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