PROPERTIES OF A PUTATIVE MUSCARINIC CHOLTNERGIC RECEPTOR FROM DROSOPHILA MELANOGASTER
N. Haim
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
Search for more papers by this authorS. Nahum
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
Search for more papers by this authorCorresponding Author
Y. Dudai
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
To whom reprint requests should be addressed.Search for more papers by this authorN. Haim
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
Search for more papers by this authorS. Nahum
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
Search for more papers by this authorCorresponding Author
Y. Dudai
Department of Neurobiology, The Weizmann Institute of Science, Rehovot, Israel
To whom reprint requests should be addressed.Search for more papers by this authorAbstract—
The powerful muscarinic antagonist [3H]quinuclidinyl benzilate (QNB) specifically binds to homogenates of Drosophila melanogaster head at a level of 65 ± 6 fmol/mg protein, with an apparent dissociation constant of 0.15–0.7 nM. The half-life of the ligand-receptor complex at 25°C is 30–40 min. Binding is inhibited by low concentrations of muscarinic ligands but not by low concentrations of nicotinic ligands, anticholinesterases or non-cholinergic drugs. Binding-sites are membrane bound and are inactivated by trypsin and by Triton X-100. Part of the activity (<20%) is released into a high speed supernatant by 2 M-NaCI. The gene coding for the putative muscarinic receptor in Drosophila is apparently not located adjacent to the gene for acetylcholinesterase
Abbreviations used:
-
- [3H]QNB
-
- [3H]quinuclidinyl benzilate
REFERENCES
- Aziz, S. A. & Eldefrawi, M. E. (1973) Cholinergic receptors of the central nervous system of insects. Pest. Biochem. Physiol. 3, 168–174.
- Bartfai, T., Anner, J., Schultzberg, M. & Mantelius, J. (1974) Partial purification and characterization of a muscarinic acetylcholine receptor from rat cerebral cortex. Biochem. biophys. Res. Commun. 59. 725–733.
- Birdsall, N. J. M. & Hulmi, E. C. (1976) Biochemical studies on muscarinic acetylcholine receptors, J. Neurochem. 27, 7–16.
- Ben-Bakak, J. & Dudai, Y. (1978) Cholinergic binding sites in rat hippocampal formation: properties and ontogenesis. Brain Res., in press.
- Changeux, J.-P., Kasai, M. & Lee, C.-Y. (1970) Use of a snake venom toxin to characterize the cholinergic receptor protein. Proc. natn. Acad. Sci. U.S.A. 67, 1241–1247.
- Corbett, J. R. (1974) The Biochemical Modce of Action of Pesticides. Academic Press, London .
- Cuatrecasas, P. & Hollenberg, M. D. (1976) Membrane receptors and hormone action. Adv. Protein Chem. 30, 251–451.
- Donnellan, J. F., Jewess, P. J. & Cattell, K. J. (1975) Subcellular localization and properties of a cholinergic receptor isolated from housefly heads. J. Neurochem. 25, 623–629.
- Dudai, Y. (1977) Demonstration of an α-bungarotoxin-binding nicotinic receptor in flies. FEBS Lett. 76, 211–213.
- Dudai, Y. (1978) Properties of an α-bungarotoxin-binding cholinergic nicotinic receptor from Drosophila melanogaster. Biochem. biophys. Acta 539, 505–517.
- Dudai, Y. & Amsterdam, A. (1977) Nicotinic receptors in the brain of Drosophila melanogaster demonstrated by autoradiography with [125 I]α-bungarotoxin. Brain Res. 130, 551–555.
- Dudai, Y. & Ben-Barak, J. (1977) Muscarinic receptor in Drosophila melanogaster demonstrated by binding of[3H]quinuclidinyl benzilate. FEBS Lett. 81, 134–136.
- Eldefrawi, A. & O'brjen, R. D. (1970) Binding of muscarone by extracts of housefly brain: relationship to receptors for acetylcholine. J. Neurochem. 17, 1287–1293.
- Eldefrawi, M. E., Eldefrawi, A. T., & O'brien, R. D. (1971) Binding of five cholinergic ligands to housefly brain and Torpedo electroplax: relationship to acetylcholine receptors. Molec. Pharmac. 7, 104–110.
- Galper, J.B., Klein, W. & Catterall, W. A. (1977) Muscarinic acetylcholine receptors in developing chick heart. J. biol. Chem. 252, 8692–8699.
- Hall, J. C. & Kankel, D. R. (1976) Genetics of acetylcholinesterase in Drosophila melanogaster. Genetics, Princeton 83, 517–535.
- Jewess, P. J., Clarke, B. S. & Donnellan, J. F. (1975) Isolation of a housefly head protein fraction that exhibits high affinity binding of cholinergic ligands. Croat. Chem. Acta 47, 459–464.
- Johnson, C. D. & Russell, L. R. (1975) A rapid, simple radiometric assay for cholinesterase, suitable for multiple determinations. Analyt. Biochem. 64, 229–238.
- Kato, G., Tan, E. & Yung, J. (1972) Acetylcholinesterase, kinetic studies on the mechanism of atropine inhibition. J. biol. Chem. 247, 3186–3189.
- Kloog, Y. & Sokolovsky, M. (1978) Studies of muscarinic acetylcholine receptors from mouse brain: characterization of the interaction with antagonists. Brain Res 144, 31–48.
- Levitzki, A., Sevilia, N., Atlas, D. & Steer, M. L. (1975) Ligand specificity and characteristics of the β-adrenergic receptor in turkey erythrocyte plasma membranes. J. molec. Biol. 97, 35–53.
- Lewis, E. B. (1960) A new standard food medium. Drosohp. Inf. Serv. 34, 117–118.
- Lindsley, D. L. & Grell, E. H. (1968) Genetic variations of Drosophila melariogaster. Carnegie Institution of Washington, Publication No. 627.
- Lowry, O. H., Roseurough, N. J., Farr, A. L. & Randall, R. J. (1951) Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275.
- Mansour, N. A., Eldefrawi, M. E. & Eldefrawi, A. T. (1977) Isolation of putative acetylcholine receptors proteins from housefly brain. Biochem. 16, 4126–4132.
- Rudloff, E. (1978) Acetylcholine receptors in the central nervous system of Drosophila melanogaster. Expl. Cell Res. 111, 185–190.
- Schmidt, J. & Raftery, M. A. (1973) A simple assay for the study of solubilized acetylcholine receptors. Analyt. Biochem. 52, 349–354.
- Schmidt-Nielsen, B. K., Gepner, J. L., Teng, N. N. H. & HALL, L. M. (1977) Characterization of an α-bungarotoxin binding component from Drosophila melanogaster.J. Neurochem. 29, 1013–1029.
- Segal, M., Dudai, Y. & Amsterdam, A. (1978) Distribution of an α-bungarotoxin-binding cholinergic nicotinic receptor in rat brain. Brain Res. 148, 105–119.
- Yamamura, H. I. & Snyder, S. H. (1974) Muscarinic cholinergic binding in rat brain. Proc. natn. Acad. Sci. U.S.A. 71, 1725–1729.
