Modulation by certain conserved aspartate residues of the allosteric interaction of gallamine at the m1 muscarinic receptor
Journal of Pharmacology and Experimental Therapeutics
Muscarinic acetylcholine receptors belong to a superfamily of G-protein coupled receptors and contain within their structure several conserved aspartate residues. These residues have been implicated to play important roles in the interaction of agonists and their competitive antagonists with the receptor. In the present work, we investigated whether the same residues might also serve as important contact points for allosteric antagonists of muscarinic receptors, because the majority of these compounds are cationic in nature, or if such residues are involved in modification of receptor conformation by these antagonists. Gallamine was used as a prototype for these antagonists. Site-directed mutagenesis of the m1 muscarinic receptor subtype was utilized to define some of the molecular determinants involved in cooperative allosteric interactions. We report that substitution of the aspartate residue at position 71, but not at positions 99 and 122 with asparagine, affected the affinity of gallamine for the unliganded m1 receptor. A similar substitution at positions 71 and 99 decreased the magnitude of its cooperative effects on the binding of [3H]N- methylscopolamine. Our data suggest that these residues are implicated in cooperative interactions. At present, however, we cannot discount a more pivotal role of other residues on the receptor sequence in allosteric interactions. The data also support the notion that different molecular entities are required for the binding of allosteric antagonists as compared to the interaction of agonists and competitive antagonists at the receptor.
Lee, N., Hu, J., & El-Fakahany, E. (1992). Modulation by certain conserved aspartate residues of the allosteric interaction of gallamine at the m1 muscarinic receptor. Journal of Pharmacology and Experimental Therapeutics, 262 (1). Retrieved from https://hsrc.himmelfarb.gwu.edu/smhs_path_facpubs/1342