Rapsyn and agrin slow the metabolic degradation of the acetylcholine receptor.
Rapsyn is a 43-kDa cytoplasmic protein that clusters nicotinic acetylcholine receptors (AChR) in the postsynaptic membrane. Here we examine the effect of rapsynmediated AChR clustering on the metabolic stability of the AChR. When transfected into QT-6 fibroblasts, cell surface AChRs (alpha, beta, epsilon, and delta subunit combination) pulse labeled with 125I-alpha-bungarotoxin were degraded with a half-life of 16.4 +/- 1.1 h (mean +/- SEM). Cotransfection of rapsyn with AChR caused extensive AChR clustering and increased AChR half-life to 20.5 +/- 1.0 h. Anti-AChR antibodies such as mab 35 cause an increased AChR degradation often associated with myasthenia gravis: 80.8 +/- 2.5% of AChRs labeled at zero time were degraded over a 12-h period. Contransfection of rapsyn reduced this AChR loss to 66.4 +/- 3.8%. Rapsyn also reduced normal AChR degradation, from 53.2 +/- 2.1 to 44.2 +/- 2.2%. Muscle cell lines from wild-type myotubes displayed few AChR clusters, but treatment with neural agrin increased the number of AChR clusters 30-fold. Clustering was accompanied by reductions in AChR degradation (both in the presence and absence of mab 35) similar in magnitude to those produced by overexpression of rapsyn in QT-6 cells. In rapsyn-deficient myotubes, treatment with neural agrin neither caused AChR clustering nor reduced AChR degradation. Thus neural agrin may slow AChR degradation by inducing the rapsyn-dependent clustering of AChRs.
Phillips
W D
WD
Department of Physiology, University of Sydney, NSW, Australia.
Vladeta
D
D
Han
H
H
Noakes
P G
PG
D018171:Agrin / Q000494:pharmacology*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D005347:Fibroblasts / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D009124:Muscle Proteins / Q000235:genetics / Q000494:pharmacology*; D009132:Muscles / Q000166:cytology / Q000378:metabolism; D009154:Mutation; D011940:Receptor Aggregation / Q000187:drug effects; D011950:Receptors, Cholinergic / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000235:genetics
D018171:Agrin; D000906:Antibodies; D009124:Muscle Proteins; D011950:Receptors, Cholinergic; D011978:Receptors, Nicotinic; C051609:peripheral membrane protein 43K