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Permeability control by cholinergic receptors in Torpedo postsynaptic membranes: agonist dose-response relations measured at second and millisecond times
Abstract
A quantitative analysis of nicotine acetylcholine receptor function in Torpedo postsynaptic membranes is presented. 22Na+ efflux induced by carbamylcholine (Carb) and the partial agonist phenyltrimethylammonium (PTA) is assessed by determining dose-response relations using three approaches: (1) a filtration assay measuring responses on the 10-s time scale, (2) the same filtration assay after blocking different fractions of the receptor sites with alpha-bungarotoxin (alpha-BgTx), and (3) a rapid-mix quenched-flow technique which permits measurement of the initial rate of 22Na+ efflux on the millisecond time scale. The concentrations of agonist producing half-maximal responses in these three assays at 4 degrees C are 13, 150, and 600 microM, respectively, for Carb and 50, 50, and 200 microM, respectively, for PTA. The rate constants for 22Na+ efflux are 1.3 x 10(-4) s-1 in the absence of agonst and 65 s-1 and 0.8 s-1 in the presence of maximal concentrations of Carb and PTA, respectively, representing a stimulation of 5 x 10(5) by Carb. The Hill coefficient for the Carb response, expressed as rate constants for 22N+ efflux, is 1.97 +/- 0.06 for Carb concentrations between 3 microM and 1 mM. The inhibition of the agonist-stimulated 22Na+ efflux by alpha-BgTx is compatible with two alpha-BgTx (and acetylcholine) sites per functional unit. Inhibition of Carb responses (slow assay) by d-tubocurarine appears competitive with a KI approximately 0.5 microM, while responses to PTA are inhibited noncompetitively with KI = 0.3 microM. This paradox is due to the presence of spare receptors and to complexities in the binding of dTC to the nicotinic acetylcholine receptor. Determination of responses without the complication of spare receptors allows a meaningful comparison to direct measurements of agonist and antagonist binding in the same system. A model is proposed to account for both binding and response.
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Citations
Cited by
Acetylcholine Receptor: An Allosteric Protein
Changeux, J.-P., Devillers-Thiéry, A., & Chemouilli, P. (1984) · Science · doi:10.1126/science.6382611 · Cited by 623
Allosteric Receptors after 30 Years
Changeux, J.-P., & Edelstein, S. J. (1998) · Neuron · doi:10.1016/s0896-6273(00)80616-9 · Cited by 431
Dennis, M., Giraudat, J., Kotzyba-Hibert, F., Goeldner, M., Hirth, C., Chang, J. Y., Lazure, C., Chretien, M., & Changeux, J. P. (1988) · Biochemistry · doi:10.1021/bi00407a016 · Cited by 317
Wilson, G. G., & Karlin, A. (2001) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.98.3.1241 · Cited by 138
A kinetic mechanism for nicotinic acetylcholine receptors based on multiple allosteric transitions.
Edelstein, S. J., Schaad, O., Henry, E., Bertrand, D., & Changeux, J.-P. (1996) · Biological Cybernetics · doi:10.1007/s004220050302 · Cited by 134
Metadata
- Authors
- Neubig, R. R., & Cohen, J. B.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1980-06-10
- Volume
- 19
- Issue
- 12
- Pages
- 2770-2779
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2051482112
- Cited by
- 170
- References
- 0
- FWCI
- 9.76
- Type
- journal article
- Language
- en
- Open access
- closed
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- CoChat. (2026). Citation check: Permeability control by cholinergic receptors in Torpedo postsynaptic membranes: agonist dose-response relations measured at second and millisecond times. https://app.cochat.ai/papers/10.1021/bi00553a036
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- Neubig, R. R., & Cohen, J. B. (1980). Permeability control by cholinergic receptors in Torpedo postsynaptic membranes: agonist dose-response relations measured at second and millisecond times. Biochemistry. https://doi.org/10.1021/bi00553a036
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