Muscle-specific agrin isoforms reduce phosphorylation of AChR gamma and delta subunits in cultured muscle cells.
The accumulation of nicotinic acetylcholine receptors (AChRs) at neuromuscular synapses is triggered by agrin, a protein that is synthesized by both nerve and muscle. Nerve-derived agrin, which contains an amino acid insert at a conserved splice site in the carboxy-terminal part of the protein, induces AChR aggregation and causes tyrosine phosphorylation of the AChR beta subunit. In contrast, agrin isoforms synthesized by muscle cells lack such an insert and have no effect on AChR distribution. In order to identify possible functional roles of muscle-derived agrin we have analyzed further the effect of various fragments of recombinant agrin on AChR phosphorylation. A carboxy-terminal fragment of muscle agrin, c95A0B0, reduced AChR gamma and delta subunit phosphorylation when added to C2C12 myotubes in culture. Although c95A0B0 had no effect on AChR beta subunit phosphorylation when added alone, it inhibited AChR beta subunit phosphorylation and AChR aggregation by the nerve-specific agrin isoform c95A4B8. We conclude that muscle-derived agrin can influence, both directly and indirectly, AChR phosphorylation. Such changes may play a role in the formation, maintenance, or function of the neuromuscular junction.
Department of Physiology and Biophysics, University of Colorado Health Sciences Center, Denver, Colorado, USA.
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T
Meier
M A
MA
Ruegg
B G
BG
Wallace
D018171:Agrin / Q000235:genetics / Q000494:pharmacology* / Q000502:physiology; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D003598:Cytoskeletal Proteins / Q000378:metabolism; D049030:Dystroglycans; D007797:Laminin / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000378:metabolism; D009124:Muscle Proteins / Q000494:pharmacology* / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D011950:Receptors, Cholinergic / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology; D013329:Structure-Activity Relationship
D018171:Agrin; D003598:Cytoskeletal Proteins; D007797:Laminin; D008562:Membrane Glycoproteins; D009124:Muscle Proteins; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins; D049030:Dystroglycans