Probing the role of the cation–π interaction in the binding sites of GPCRs using unnatural amino acids
- 21 July 2009
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences of the United States of America
- Vol. 106 (29), 11919-11924
- https://doi.org/10.1073/pnas.0903260106
Abstract
We describe a general application of the nonsense suppression methodology for unnatural amino acid incorporation to probe drug-receptor interactions in functional G protein-coupled receptors (GPCRs), evaluating the binding sites of both the M2 muscarinic acetylcholine receptor and the D2 dopamine receptor. Receptors were expressed in Xenopus oocytes, and activation of a G protein-coupled, inward-rectifying K(+) channel (GIRK) provided, after optimization of conditions, a quantitative readout of receptor function. A number of aromatic amino acids thought to be near the agonist-binding site were evaluated. Incorporation of a series of fluorinated tryptophan derivatives at W6.48 of the D2 receptor establishes a cation-pi interaction between the agonist dopamine and W6.48, suggesting a reorientation of W6.48 on agonist binding, consistent with proposed "rotamer switch" models. Interestingly, no comparable cation-pi interaction was found at the aligning residue in the M2 receptor.This publication has 45 references indexed in Scilit:
- Nicotine binding to brain receptors requires a strong cation–π interactionNature, 2009
- The 2.6 Angstrom Crystal Structure of a Human A 2A Adenosine Receptor Bound to an AntagonistScience, 2008
- Structure of a β1-adrenergic G-protein-coupled receptorNature, 2008
- High-Resolution Crystal Structure of an Engineered Human β 2 -Adrenergic G Protein–Coupled ReceptorScience, 2007
- Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 1: Minimizing misacylationRNA, 2007
- In vivo incorporation of multiple unnatural amino acids through nonsense and frameshift suppressionProceedings of the National Academy of Sciences of the United States of America, 2006
- Mapping the Gβγ-binding Sites in GIRK1 and GIRK2 Subunits of the G Protein-activated K+ ChannelOnline Journal of Public Health Informatics, 2003
- Construction of a sequence motif characteristic of aminergic G protein–coupled receptorsProtein Science, 2003
- Specific Tryptophan UV-Absorbance Changes Are Probes of the Transition of Rhodopsin to Its Active StateBiochemistry, 1996
- Unconventional ionic hydrogen bonds. 2. NH+.cntdot..cntdot..cntdot..pi.. Complexes of onium ions with olefins and benzene derivativesJournal of the American Chemical Society, 1985