Incorporation of a gephyrin‐binding motif targets NMDA receptors to gephyrin‐rich domains in HEK 293 cells

The peripheral membrane protein gephyrin is essential for the postsynaptic localization of inhibitory glycine receptors (GlyRs). Binding of gephyrin to the GlyR beta subunit is mediated by a sequence motif located in the intracellular loop region connecting transmembrane segments 3 and 4. Here, insertion of this binding motif is shown to alter the subcellular distribution of an excitatory neurotransmitter receptor in transfected mammalian cells. Upon coexpression with gephyrin, a mutant N-methyl-D-aspartate (NMDA) receptor containing NMDA receptor 1 (NR1) subunits which harboured a gephyrin-binding motif within its cytoplasmic tail region, was targeted to intracellular gephyrin-rich domains, as previously observed for the GlyR beta subunit. Our data indicate that a gephyrin-binding motif located in a cytoplasmic domain of an integral membrane protein suffices for routing to gephyrin-rich domains.