Lyn Dissociation from Phosphorylated FcεRI Subunits: A New Regulatory Step in the FcεRI Signaling Cascade Revealed by Studies of FcεRI Dimer Signaling Activity

Abstract

AbstractCross-linking the heterotrimeric (αβγ2) IgE receptor, FcεRI, of mast cells activates two tyrosine kinases: Lyn, which phosphorylates β and γ subunit immunoreceptor tyrosine-based activation motifs, and Syk, which binds γ-phospho-immunoreceptor tyrosine-based activation motifs and initiates cellular responses. We studied three FcεRI-dimerizing mAbs that maintain similar dispersed distributions over the surface of RBL-2H3 mast cells but elicit very different signaling responses. Specifically, mAb H10 receptor dimers induce very little inositol 1,4,5-trisphosphate synthesis, Ca2+ mobilization, secretion, spreading, ruffling, and actin plaque assembly, whereas dimers generated with the other anti-FcεRI mAbs induce responses that are only modestly lower than that to multivalent Ag. H10 receptor dimers activate Lyn and support FcεRI β and γ subunit phosphorylation but are poor Syk activators compared with Ag and the other anti-FcεRI mAbs. H10 receptor dimers have two other distinguishing features. First, they induce stable complexes between activated Lyn and receptor subunits. Second, the predominant Lyn-binding phospho-β isoform found in mAb H10-treated cells is a less tyrosine phosphorylated, more electrophoretically mobile species than the predominant isoform in Ag-treated cells that does not coprecipitate with Lyn. These studies implicate Lyn dissociation from highly phosphorylated receptor subunits as a new regulatory step in the FcεRI signaling cascade required for Syk activation and signal progression.