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Association of acetylcholine receptors with peripheral membrane proteins: evidence from antibody-induced coaggregation

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Abstract

Acetylcholine receptors (AChR) are associated with several peripheral membrane proteins that are concentrated on the cytoplasmic face of the plasma membrane at the neuromuscular junction, and at aggregates of AChR that form in vitro. We tested the linkage among these proteins by inducing microaggregation of AChR, then determining if a given peripheral membrane protein accumulated with the receptors in microaggregates. In most experiments, we used isolated membrane fragments that are rich in AChR and accessible to antibodies against intracellular antigens. We showed that the 43 kD receptor-associated protein always aggregated with AChR, whether microaggregation was driven by antibodies to the 43 kD protein, or to the receptor itself. Antibodies to the 58 kD receptor-associated protein also always aggregated the 58 kD protein with the receptor. Our results are consistent with a model for AChR-rich membrane in which the 43 kD and 58 kD proteins are both closely associated with the AChR.

When we induced microaggregation in intact muscle cells with anti-AChR antibodies, our results were less definitive. The 43 kD receptor-associated protein microaggregated with AChR, but the 58 kD protein was not especially enriched at AChR microaggregates. We discuss the advantages of using isolated AChR-rich membrane fragments to study the association of AChR with peripheral membrane proteins.

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Authors and Affiliations

  1. Department of Physiology, University of Maryland School of Medicine, 660 W. Redwood St., 21201, Baltimore, Maryland

    R. J. Bloch & P. W. Luther

  2. Department of Anatomy, University of Maryland School of Medicine, Baltimore, Maryland

    D. W. Pumplin

  3. Department of Physiology, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA

    R. Sealock & S. C. Froehner

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  1. R. J. Bloch
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  2. R. Sealock
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  3. D. W. Pumplin
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  4. P. W. Luther
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  5. S. C. Froehner
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Additional information

We thank J. Strong for his expertise in obtaining the data for Figs. 6–8, and W. Resneck, A. O'Neill, and K. Douville for their assistance throughout this work. Our research has been supported by grants from the Muscular Dystrophy Association (to R.J.B., R.S., D.W.P. and S.C.F.) and from the National Institutes of Health (NS17282 to R.J.B.; NS27171 to P.W.L.; NS15513 to D.W.P.; NS15293 to R.S.; NS14871 to S.C.F.).

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Bloch, R.J., Sealock, R., Pumplin, D.W. et al. Association of acetylcholine receptors with peripheral membrane proteins: evidence from antibody-induced coaggregation. J. Membarin Biol. 138, 13–28 (1994). https://doi.org/10.1007/BF00211065

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  • DOI: https://doi.org/10.1007/BF00211065

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