Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor
The positive allosteric modulators (PAMs) of the α7 nicotinic receptor <i>N</i>-(5-Cl-2-hydroxyphenyl)-<i>N</i>′-[2-Cl-5-(trifluoromethyl)phenyl]-urea (NS-1738) and (<i>E</i>)-3-(furan-2-yl)-<i>N</i>-(<i>p</i>-tolyl)-acrylamide (PAM-2) pote...
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| Format: | Article |
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MDPI AG
2023-04-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/13/4/698 |
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| author | Spencer R. Pierce Allison L. Germann Sophia Q. Xu Saumith L. Menon Marcelo O. Ortells Hugo R. Arias Gustav Akk |
| author_facet | Spencer R. Pierce Allison L. Germann Sophia Q. Xu Saumith L. Menon Marcelo O. Ortells Hugo R. Arias Gustav Akk |
| author_sort | Spencer R. Pierce |
| collection | DOAJ |
| description | The positive allosteric modulators (PAMs) of the α7 nicotinic receptor <i>N</i>-(5-Cl-2-hydroxyphenyl)-<i>N</i>′-[2-Cl-5-(trifluoromethyl)phenyl]-urea (NS-1738) and (<i>E</i>)-3-(furan-2-yl)-<i>N</i>-(<i>p</i>-tolyl)-acrylamide (PAM-2) potentiate the α1β2γ2L GABA<sub>A</sub> receptor through interactions with the classic anesthetic binding sites located at intersubunit interfaces in the transmembrane domain of the receptor. In the present study, we employed mutational analysis to investigate in detail the involvement and contributions made by the individual intersubunit interfaces to receptor modulation by NS-1738 and PAM-2. We show that mutations to each of the anesthetic-binding intersubunit interfaces (β+/α−, α+/β−, and γ+/β−), as well as the orphan α+/γ− interface, modify receptor potentiation by NS-1738 and PAM-2. Furthermore, mutations to any single interface can fully abolish potentiation by the α7-PAMs. The findings are discussed in the context of energetic additivity and interactions between the individual binding sites. |
| first_indexed | 2024-03-11T05:12:04Z |
| format | Article |
| id | doaj.art-7b45852e7ea54e64b1ae9c0efcc1c93d |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-11T05:12:04Z |
| publishDate | 2023-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-7b45852e7ea54e64b1ae9c0efcc1c93d2023-11-17T18:30:04ZengMDPI AGBiomolecules2218-273X2023-04-0113469810.3390/biom13040698Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic ReceptorSpencer R. Pierce0Allison L. Germann1Sophia Q. Xu2Saumith L. Menon3Marcelo O. Ortells4Hugo R. Arias5Gustav Akk6Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USADepartment of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USADepartment of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USADepartment of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USAFacultad de Medicina, Universidad de Morón, CONICET, Morón 1708, ArgentinaDepartment of Pharmacology and Physiology, Oklahoma State University College of Osteopathic Medicine, Tahlequah, OK 74464, USADepartment of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, USAThe positive allosteric modulators (PAMs) of the α7 nicotinic receptor <i>N</i>-(5-Cl-2-hydroxyphenyl)-<i>N</i>′-[2-Cl-5-(trifluoromethyl)phenyl]-urea (NS-1738) and (<i>E</i>)-3-(furan-2-yl)-<i>N</i>-(<i>p</i>-tolyl)-acrylamide (PAM-2) potentiate the α1β2γ2L GABA<sub>A</sub> receptor through interactions with the classic anesthetic binding sites located at intersubunit interfaces in the transmembrane domain of the receptor. In the present study, we employed mutational analysis to investigate in detail the involvement and contributions made by the individual intersubunit interfaces to receptor modulation by NS-1738 and PAM-2. We show that mutations to each of the anesthetic-binding intersubunit interfaces (β+/α−, α+/β−, and γ+/β−), as well as the orphan α+/γ− interface, modify receptor potentiation by NS-1738 and PAM-2. Furthermore, mutations to any single interface can fully abolish potentiation by the α7-PAMs. The findings are discussed in the context of energetic additivity and interactions between the individual binding sites.https://www.mdpi.com/2218-273X/13/4/698GABA<sub>A</sub> receptorα7 nicotinic receptormodulationpotentiatorallostery |
| spellingShingle | Spencer R. Pierce Allison L. Germann Sophia Q. Xu Saumith L. Menon Marcelo O. Ortells Hugo R. Arias Gustav Akk Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor Biomolecules GABA<sub>A</sub> receptor α7 nicotinic receptor modulation potentiator allostery |
| title | Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor |
| title_full | Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor |
| title_fullStr | Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor |
| title_full_unstemmed | Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor |
| title_short | Mutational Analysis of Anesthetic Binding Sites and Their Effects on GABA<sub>A</sub> Receptor Activation and Modulation by Positive Allosteric Modulators of the α7 Nicotinic Receptor |
| title_sort | mutational analysis of anesthetic binding sites and their effects on gaba sub a sub receptor activation and modulation by positive allosteric modulators of the α7 nicotinic receptor |
| topic | GABA<sub>A</sub> receptor α7 nicotinic receptor modulation potentiator allostery |
| url | https://www.mdpi.com/2218-273X/13/4/698 |
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