During antigen recognition by T cells, CD4 and the T-cell receptor (TCR)/CD3/zeta complex are thought to interact with the same major histocompatibility complex II molecule in a stable ternary complex. Evidence has suggested that the association of CD4 with TCR/CD3/zeta requires the interaction of the protein tyrosine kinase p56lck with CD4. We have taken a biochemical approach to understand the mechanism by which p56lck and, in particular, its src homology (SH) 2 domain contributes to the association of CD4 with TCR/CD3/zeta during activation. We have previously shown that the p56lck SH2 domain binds directly to tyrosine-phosphorylated ZAP-70. Here we formally demonstrate the in vivo association of p56lck with the homologous protein tyrosine kinases Syk and ZAP-70 after CD3 stimulation of Jurkat cells. A tyrosine-phosphorylated peptide containing the sequence predicted to be optimal for binding to the SH2 domain of src family kinases specifically competes for this association, indicating that tyrosine-phosphorylated ZAP-70 and Syk bind to p56lck by an SH2-mediated interaction. We also show that the same peptide is able to compete for the activation-dependent TCR/CD4 association in Jurkat cells. Moreover, ZAP-70 and CD4 cocap only after CD3 stimulation in human T lymphoblasts. We propose that the interaction of the p56lck SH2 domain with zeta-associated tyrosine-phosphorylated ZAP-70 and/or Syk enables CD4 to associate with antigen-stimulated TCR/CD3/zeta complexes.
Article|
June 01 1995
Syk and ZAP-70 mediate recruitment of p56lck/CD4 to the activated T cell receptor/CD3/zeta complex.
M Thome,
M Thome
Department of Immunology, Institut Pasteur, Paris, France.
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P Duplay,
P Duplay
Department of Immunology, Institut Pasteur, Paris, France.
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M Guttinger,
M Guttinger
Department of Immunology, Institut Pasteur, Paris, France.
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O Acuto
O Acuto
Department of Immunology, Institut Pasteur, Paris, France.
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M Thome
Department of Immunology, Institut Pasteur, Paris, France.
P Duplay
Department of Immunology, Institut Pasteur, Paris, France.
M Guttinger
Department of Immunology, Institut Pasteur, Paris, France.
O Acuto
Department of Immunology, Institut Pasteur, Paris, France.
Online ISSN: 1540-9538
Print ISSN: 0022-1007
J Exp Med (1995) 181 (6): 1997–2006.
Citation
M Thome, P Duplay, M Guttinger, O Acuto; Syk and ZAP-70 mediate recruitment of p56lck/CD4 to the activated T cell receptor/CD3/zeta complex.. J Exp Med 1 June 1995; 181 (6): 1997–2006. doi: https://doi.org/10.1084/jem.181.6.1997
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