Peptide extraction by alkaline treatment is accompanied by rearrangement of the membrane‐bound acetylcholine receptor from <i>torpedo marmorata</i>

Barrantes, Francisco J.; Neugebauer, D.-Ch.; Zingsheim, H. P.

doi:10.1016/0014-5793(80)80131-1

Cited by 109 publications

(61 citation statements)

References 18 publications

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“…In clusters that occur on the ventral surfaces of rat myotubes, for example, vinculin is concentrated in membrane areas that interdigitate with regions of high AChR density (6). On the other hand, there is now biochemical (9) and ultrastructural (44,45) evidence that the 43K protein is closely associated with and possibly linked noncovalently to AChR in Torpedo postsynaptic membranes, an interaction that may restrict the mobility of the receptors (4,25). Thus, the 43K protein is a good candidate for the mediation of the interaction between the receptors and the cytoskeleton.…”

Section: Discussionmentioning

confidence: 99%

“…Exposure to alkaline pH causes dissociation of this protein from the membranes (26) with a concomitant increase in the rotational and translational mobility of the receptor (4,10,25,41). The 43K protein is distinct from actin (36) and creatine kinase (5,16) and lies on the cytoplasmic side of the membrane (27,36,45,47,49) in close enough proximity to the receptor so that chemical crosslinking between the two proteins can be induced with a bifunctional reagent (9).…”

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confidence: 99%

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Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Peng¹,

Froehner²

1985

The Journal of Cell Biology

107

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The postsynaptic membrane from Torpedo electric organ contains, in addition to the acetylcholine receptor (AChR), a major peripheral membrane protein of ~43,000 mol wt (43K protein). Previous studies have shown that this protein is closely associated with AChR and may be involved in anchoring receptors to the postsynaptic membrane. In this study, binding sites for monoclonal antibodies (mabs) to the 43K protein have been compared to the distribution of AChR in Xenopus laevis muscle cells in culture. In double label immunofluorescence experiments, clusters of AChR that occur spontaneously on these cells were stained with anti-43K mabs. Newly formed receptor clusters induced with positive polypeptidecoated latex beads were also stained with anti-43K mabs as early as 12 h after the application of the beads. Exact correspondence in the distribution of the anti-43K protein binding sites and the AChR was found in both types of clusters. These results suggest that the 43K protein becomes associated with AChR clusters during a period of active postsynaptic membrane differentiation. Thus, this protein may participate in the clustering process.An early event during the formation of the neuromuscular synapse is the aggregation of acetylcholine receptors (AChR) ~ at the postsynaptic membrane in response to innervation (1,15,50). Tissue cultures of skeletal muscle cells are an invaluable tool for the study of this process. In addition to neurally induced receptor aggregates, AChR also form clusters spontaneously (the "hot spots"; 2, 14, 48) or in response to exogenously applied stimuli in tissue culture (l 1, 23, 31, 34, 42). Although these extrasynaptic AChR clusters have proven to be a convenient system to study, the cellular and molecular mechanisms involved in the clustering process remain largely unknown. Recent studies have indicated that the cytoskeleton may be involved in the aggregation of AChR (12,28,38), but the precise nature of the AChR-cytoskeleton interaction remains elusive.Highly purified postsynaptic membranes from Torpedo Abbreviations used in this paper: AChR, acetylcholine receptor(s); FITC, fluorescein isothiocyanate; 43K protein, the major membranebound 43,000-mol-wt protein isolated from Torpedo postsynaptic membrane; mab, monoclonal antibody; R-BTX, tetramethyl rhodamine-conjugated a-bungarotoxin. 1698electric organ contain, in addition to the ACh receptor', a major peripheral membrane protein of-43,000 mol wt (43K protein) (13,19,26,36,46). Exposure to alkaline pH causes dissociation of this protein from the membranes (26) with a concomitant increase in the rotational and translational mobility of the receptor (4, 10, 25, 41). The 43K protein is distinct from actin (36) and creatine kinase (5, 16) and lies on the cytoplasmic side of the membrane (27,36,45,47,49) in close enough proximity to the receptor so that chemical crosslinking between the two proteins can be induced with a bifunctional reagent (9). Recent results indicate that the 43K protein interacts in a hydrophobic manner with th...

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Peng¹,

Froehner²

1985

The Journal of Cell Biology

107

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“…The 43 kDa protein is not required for ligand-gated ion flux through the receptor channel [9]. However, its removal from both Torpedo [10,11] and rat postsynaptic membranes [8] by alkaline extraction is accompanied by increased mobility and rearrangement of the receptors. Thus, although its function is not known, the 43 kDa protein is thought to play some role in anchoring or stabilizing AChR at synaptic sites (review [13]).…”

Section: Introductionmentioning

confidence: 99%

Expression of RNA transcripts for the postsynaptic 43 kDa protein in innervated and denervated rat skeletal muscle

Froehner

1989

FEBS Letters

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A cDNA clone encoding the mouse muscle postsynaptic 43 kDa protein was isolated and sequenced. The amino acid sequence of this protein, which is closely associated with nicotinic acetylcholine receptors at Torpedo electrocyte and vertebrate skeletal muscle synapses, is very similar in different species. A cysteine-rich region homologous to part of the regulatory domain of protein kinase C may be important in interactions of this protein with the lipid bilayer. RNA transcripts for the 43 kDa protein increase only 2-3-fold after denervation of rat skeletal muscle, in sharp contrast to the a-subunit of the muscle nicotinic receptor which increases more than 30-fold. Thus, the expression of these two proteins is regulated by different mechanisms.

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“…The 43 k polypeptide is a peripheral protein which is released from the subsynaptic membrane by treatment at pH 11. Its alkaline extraction does not significantly change the known regulatory properties of the ACh receptor protein [ 13-161 but alters its thermal stability [ 171 and its rotational [ 18-211 and translational [22] diffusion. Most likely, the 43 k protein plays a 'structural' role in maintaining the ACh receptor protein in a highly immobilized and stable aggregated form in the subsynaptic membrane.…”

Section: Introductionmentioning

confidence: 99%

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[³H]trimethisoquin

et al. 1980

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Peptide extraction by alkaline treatment is accompanied by rearrangement of the membrane‐bound acetylcholine receptor from torpedo marmorata

Cited by 109 publications

References 18 publications

Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Expression of RNA transcripts for the postsynaptic 43 kDa protein in innervated and denervated rat skeletal muscle

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[³H]trimethisoquin

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Peptide extraction by alkaline treatment is accompanied by rearrangement of the membrane‐bound acetylcholine receptor from torpedo marmorata

Cited by 109 publications

References 18 publications

Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Association of the postsynaptic 43K protein with newly formed acetylcholine receptor clusters in cultured muscle cells.

Expression of RNA transcripts for the postsynaptic 43 kDa protein in innervated and denervated rat skeletal muscle

Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[3H]trimethisoquin

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Conditions for the selective labelling of the 66 000 dalton chain of the acetylcholine receptor by the covalent non‐competitive blocker 5‐azido‐[³H]trimethisoquin