Paper Citing NAMD - Abstract
Abdul-Ridha, Alaa; Lopez, Laura; Keov, Peter; Thal, David M.; Mistry, Shailesh N.; Sexton, Patrick M.; Lane, J. Robert; Canals, Meritxell; Christopoulos, Arthur
Molecular Determinants of Allosteric Modulation at the M-1 Muscarinic Acetylcholine Receptor
JOURNAL OF BIOLOGICAL CHEMISTRY, 289:6067-6079, FEB 28 2014
Background: BQCA is a selective allosteric modulator of the M-1 mAChR. Results: Residues that govern BQCA activity were identified using mutagenesis and molecular modeling. Conclusion: BQCA likely occupies a pocket overlapping prototypical mAChR modulators and gains selectivity through cooperativity with orthosteric ligands. Significance: Understanding the structural basis of BQCA function can provide insight into the design of more tailored allosteric ligands. Benzylquinolone carboxylic acid (BQCA) is an unprecedented example of a selective positive allosteric modulator of acetylcholine at the M-1 muscarinic acetylcholine receptor (mAChR). To probe the structural basis underlying its selectivity, we utilized site-directed mutagenesis, analytical modeling, and molecular dynamics to delineate regions of the M-1 mAChR that govern modulator binding and transmission of cooperativity. We identified Tyr-85(2.64) in transmembrane domain 2 (TMII), Tyr-179 and Phe-182 in the second extracellular loop (ECL2), and Glu-397(7.32) and Trp-400(7.35) in TMVII as residues that contribute to the BQCA binding pocket at the M-1 mAChR, as well as to the transmission of cooperativity with the orthosteric agonist carbachol. As such, the BQCA binding pocket partially overlaps with the previously described common allosteric site in the extracellular vestibule of the M-1 mAChR, suggesting that its high subtype selectivity derives from either additional contacts outside this region or through a subtype-specific cooperativity mechanism. Mutation of amino acid residues that form the orthosteric binding pocket caused a loss of carbachol response that could be rescued by BQCA. Two of these residues (Leu-102(3.29) and Asp-105(3.32)) were also identified as indirect contributors to the binding affinity of the modulator. This new insight into the structural basis of binding and function of BQCA can guide the design of new allosteric ligands with tailored pharmacological properties.
