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• W151716105 abstract "Research Article1 January 1993free access Functional role of proline and tryptophan residues highly conserved among G protein-coupled receptors studied by mutational analysis of the m3 muscarinic receptor. J. Wess J. Wess National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author S. Nanavati S. Nanavati National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author Z. Vogel Z. Vogel National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author R. Maggio R. Maggio National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author J. Wess J. Wess National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author S. Nanavati S. Nanavati National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author Z. Vogel Z. Vogel National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author R. Maggio R. Maggio National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. Search for more papers by this author Author Information J. Wess1, S. Nanavati1, Z. Vogel1 and R. Maggio1 1National Institute of Neurological Disorders and Stroke, Laboratory of Molecular Biology, Bethesda, MD 20892. The EMBO Journal (1993)12:331-338https://doi.org/10.1002/j.1460-2075.1993.tb05661.x PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Most G protein-coupled receptors contain a series of highly conserved proline and tryptophan residues within their hydrophobic transmembrane domains (TMD I-VII). To study their potential role in ligand binding and receptor function, the rat m3 muscarinic acetylcholine receptor was used as a model system. A series of mutant receptors in which the conserved proline and tryptophan residues were individually replaced with alanine and phenylalanine, respectively, was created and transiently expressed in COS-7 cells. [3H]N-methylscopolamine ([3H]NMS) saturation binding studies showed that three of the seven mutant receptors studied (Pro242-->Ala, TMD V; Pro505-->Ala, TMD VI; Pro540-->Ala, TMD VII) were expressed at 35–100 times lower levels than the wild-type receptor while displaying ‘m3-like’ antagonist binding affinities. Pro201-->Ala (TMD IV) showed drastically reduced binding affinities (up to 450-fold) for both muscarinic agonists and antagonists. Whereas most mutant receptors retained strong functional activity, Pro540-->Ala (TMD VII) was found to be severely impaired in its ability to stimulate carbachol-induced phosphatidyl inositol hydrolysis (Emax approximately 25% of wild type m3). Interestingly, this mutant receptor bound muscarinic agonists with 7- to 19-fold higher affinities than the wild type receptor. The Trp-->Phe substitutions (Trp192-->Phe, TMD IV; Trp503-->Phe, TMD VI; Trp530-->Phe, TMD VII) resulted in less pronounced changes (compared with the Pro-->Ala mutant receptors) in both ligand binding and receptor function. Our data indicate that the proline residues that are highly conserved across the entire superfamily of G protein-coupled receptors play key roles in receptor expression, ligand binding and receptor activation. Previous ArticleNext Article Volume 12Issue 11 January 1993In this issue RelatedDetailsLoading ..." @default.
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• W151716105 title "Functional role of proline and tryptophan residues highly conserved among G protein-coupled receptors studied by mutational analysis of the m3 muscarinic receptor." @default.
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