Evidence for in Situ and in VitroAssociation between β-Dystroglycan and the Subsynaptic 43K Rapsyn Protein

Abstract

The accumulation of dystrophin and associated proteins at the postsynaptic membrane of the neuromuscular junction and their co-distribution with nicotinic acetylcholine receptor (AChR) clusters in vitro suggested a role for the dystrophin complex in synaptogenesis. Co-transfection experiments in which alpha- and beta-dystroglycan form a complex with AChR and rapsyn, a peripheral protein required for AChR clustering (Apel, D. A., Roberds, S. L., Campbell, K. P., and Merlie, J. P. (1995) Neuron 15, 115-126), suggested that rapsyn functions as a link between AChR and the dystrophin complex. We have investigated the interaction between rapsyn and beta-dystroglycan in Torpedo AChR-rich membranes using in situ and in vitro approaches. Cross-linking experiments were carried out to study the topography of postsynaptic membrane polypeptides. A cross-linked product of 90 kDa was labeled by antibodies to rapsyn and beta-dystroglycan; this demonstrates that these polypeptides are in close proximity to one another. Affinity chromatography experiments and ligand blot assays using rapsyn solubilized from Torpedo AChR-rich membranes and constructs containing beta-dystroglycan C-terminal fragments show that a rapsyn-binding site is present in the juxtamembranous region of the cytoplasmic tail of beta-dystroglycan. These data point out that rapsyn and dystroglycan interact in the postsynaptic membrane and thus reinforce the notion that dystroglycan could be involved in synaptogenesis.