Tyrosine phosphorylation of nicotinic acetylcholine receptor mediates Grb2 binding.
Tyrosine phosphorylation of the nicotinic acetylcholine receptor (AChR) is associated with an altered rate of receptor desensitization and also may play a role in agrin-induced receptor clustering. We have demonstrated a previously unsuspected interaction between Torpedo AChR and the adaptor protein Grb2. The binding is mediated by the Src homology 2 (SH2) domain of Grb2 and the tyrosine-phosphorylated delta subunit of the AChR. Dephosphorylation of the delta subunit abolishes Grb2 binding. A cytoplasmic domain of the delta subunit contains a binding motif (pYXNX) for the SH2 domain of Grb2. Indeed, a phosphopeptide corresponding to this region of the delta subunit binds to Grb2 SH2 fusion proteins with relatively high affinity, whereas a peptide lacking phosphorylation on tyrosine exhibits no binding. Grb2 is colocalized with the AChR on the innervated face of Torpedo electrocytes. Furthermore, Grb2 specifically copurifies with AChR solubilized from postsynaptic membranes. These data suggest a novel role for tyrosine phosphorylation of the AChR in the initiation of a Grb2-mediated signaling cascade at the postsynaptic membrane.
Colledge
M
M
Department of Physiology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Froehner
S C
SC
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D004557:Electric Organ / Q000166:cytology / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D010748:Phosphopeptides / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013570:Synaptic Membranes / Q000378:metabolism; D014018:Tissue Distribution; D014101:Torpedo; D014443:Tyrosine / Q000378:metabolism*; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D051380:GRB2 Adaptor Protein; D010748:Phosphopeptides; D011506:Proteins; D011978:Receptors, Nicotinic; D011993:Recombinant Fusion Proteins; D019000:Phosphotyrosine; D014443:Tyrosine