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Keywords:

  • Noncovalent complexes;
  • CD3;
  • Tyrosine kinases;
  • Glycosylphosphatidylinositol

Abstract

A number of T cell surface transmembrane molecules such as CD2, CD4, CD8, lymphocyte functional antigen (LFA)-1 and CD45 are known to interact functionally with the T cell receptor (TCR) complex during T cell activation. Several previous communications have also reported physical associations between some of these molecules. On the other hand, there are indications that signaling through T cell surface molecules anchored via glycosylphosphatidylinositol (GPI), such as Thy-1, Ly-6 or CD59, is dependent on the TCR. Therefore, it was of interest to determine in a systematic way which T cell surface molecules are noncovalently associated with the TCR/CD3 complex and with the major intracellular signaling molecules, the protein tyrosine kinases of the Src family. To this aim, membrane proteins of human thymoma HPB-ALL cells were solubilized in a solution of the mild detergent Brij-58 and subjected to immunoprecipitation followed by in vitro kinase assays. Two types of large complexes containing protein tyrosine kinases were observed: the first one contained CD3 and the transmembrane proteins CD2, CD4, CD5, CD6, CD7, CD8, CD11a, CD38, CD43, CD45, CD71, CD98 and CD99 and the other contained mainly the GPI-anchored proteins CD48, CD55, CD59 and CDw108 as well as a fraction of CD4 and CD8. The GPI-anchored protein complexes were of larger size and lower buoyant density than the CD3 complexes. In agreement with these biochemical data, co-cross-linking of CD3 with most of the relevant transmembrane proteins on the surface of another T cell line, Jurkat, markedly enhanced tyrosine phosphorylation of several intracellular proteins. These data indicate the existence of at least two types of membrane microdomains of very different composition in the membranes of T cells which may play a role in signaling through different types of receptors and in functional cooperation between TCR/CD3 and various accessory molecules.