Down-regulation of Protein-tyrosine Phosphatases Activates an Immune Receptor in the Absence of Its Translocation into Lipid Rafts

The earliest known biochemical step that occurs after ligand binding to the multichain immune recognition receptor is tyrosine phosphorylation of the receptor subunits. In mast cells and basophils activated by multivalent antigen-IgE complexes, this step is mediated by Src family kinase Lyn, which phosphorylates the high affinity IgE receptor (Fc epsilon RI). However, the exact molecular mechanism of this phosphorylation step is incompletely understood. In this study, we tested the hypothesis that changes in activity and/ or topography of protein-tyrosine phosphatases (PTPs) could play a major role in the Fc epsilon RI triggering. We found that exposure of rat basophilic leukemia cells or mouse bone marrow-derived mast cells to PTP inhibitors, H2O2 or pervanadate, induced phosphorylation of the Fc epsilon RI subunits, similarly as Fc epsilon RI triggering. Interestingly, and in sharp contrast to antigen-induced activation, neither H2O2 nor pervanadate induced any changes in the association of Fc epsilon RI with detergent-resistant membranes and in the topography of Fc epsilon RI detectable by electron microscopy on isolated plasma membrane sheets. In cells stimulated with pervanadate, H2O2 or antigen, enhanced oxidation of active site cysteine of several PTPs was detected. Unexpectedly, most of oxidized phosphatases bound to the plasma membrane were associated with the actin cytoskeleton. Several PTPs (SHP-1, SHP-2, hematopoietic PTP, and PTP-MEG2) showed changes in their enzymatic activity and/or oxidation state during activation. Based on these and other data, we propose that down-regulation of enzymatic activity of PTPs and/or changes in their accessibility to the substrates play a key role in initial tyrosine phosphorylation of the Fc epsilon RI and other multichain immune receptors.