Identification of phosphorylation sites on AChR δ-subunit associated with dispersal of AChR clusters on the surface of muscle cells

Anjaruwee S. Nimnual, Weise Chang, Nan Shan Chang, Anthony F. Ross, Marina S. Gelman, Joav M. Prives

Research output: Contribution to journalArticlepeer-review

24 Citations (Scopus)

Abstract

The innervation of embryonic skeletal muscle cells is marked by the redistribution of nicotinic acetylcholine receptors (AChRs) on muscle surface membranes into high-density patches at nerve-muscle contacts. To investigate the role of protein phosphorylation pathways in the regulation of AChR surface distribution, we have identified the sites on AChR δ-subunits that undergo phosphorylation associated with AChR cluster dispersal in cultured myotubes. We found that PKC-catalyzed AChR phosphorylation is targeted to Ser378, Ser393, and Ser450, all located in the major intracellular domain of the AChR δ-subunit. Adjacent to one of these sites is a PKA consensus target site (Ser377) that was efficiently phosphorylated by purified PKA in vitro. The PKC activator 12-O-tetradecanoylphorbol-13- acetate (TPA) and the phosphoprotein phosphatase inhibitor okadaic acid (OA) produced increased phosphorylation of AChR δ-subunits on the three serine residues that were phosphorylated by purified PKC in vitro. In contrast, treatment of these cells with the PKA activator forskolin, or with the cell- permeable cAMP analogue 8-bromo-cAMP, did not alter the phosphorylation state of surface AChR, suggesting that PKA does not actively phosphorylate the δ- subunit in intact chick myotubes. The effects of TPA and OA included an increase in the proportion of surface AChR that is extracted in Triton X- 100, as well as the spreading of AChR from cluster regions to adjacent areas of the muscle cell surface. These findings suggest that PKC-catalyzed phosphorylation on the identified serine residues of AChR δ-subunits may play a role in the surface distribution of these receptors.

Original languageEnglish
Pages (from-to)14823-14832
Number of pages10
JournalBiochemistry
Volume37
Issue number42
DOIs
Publication statusPublished - 1998 Oct 20

All Science Journal Classification (ASJC) codes

  • Biochemistry

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