Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations
Abstract Mutations that constitutively activate G protein-coupled receptors (GPCRs), known as constitutively activating mutations (CAMs), modify cell signaling and interfere with drugs, resulting in diseases with limited treatment options. We utilize fluorescence imaging at the single-molecule level...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-11-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-023-05603-6 |
| _version_ | 1797414264839864320 |
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| author | Shushu Wei Niloofar Gopal Pour Sriram Tiruvadi-Krishnan Arka Prabha Ray Naveen Thakur Matthew T. Eddy Rajan Lamichhane |
| author_facet | Shushu Wei Niloofar Gopal Pour Sriram Tiruvadi-Krishnan Arka Prabha Ray Naveen Thakur Matthew T. Eddy Rajan Lamichhane |
| author_sort | Shushu Wei |
| collection | DOAJ |
| description | Abstract Mutations that constitutively activate G protein-coupled receptors (GPCRs), known as constitutively activating mutations (CAMs), modify cell signaling and interfere with drugs, resulting in diseases with limited treatment options. We utilize fluorescence imaging at the single-molecule level to visualize the dynamic process of CAM-mediated activation of the human A2A adenosine receptor (A2AAR) in real time. We observe an active-state population for all CAMs without agonist stimulation. Importantly, activating mutations significantly increase the population of an intermediate state crucial for receptor activation, notably distinct from the addition of a partner G protein. Activation kinetics show that while CAMs increase the frequency of transitions to the intermediate state, mutations altering sodium sensitivity increase transitions away from it. These findings indicate changes in GPCR function caused by mutations may be predicted based on whether they favor or disfavor formation of an intermediate state, providing a framework for designing receptors with altered functions or therapies that target intermediate states. |
| first_indexed | 2024-03-09T05:30:31Z |
| format | Article |
| id | doaj.art-a5791267b9534817a20ef741b4663262 |
| institution | Directory Open Access Journal |
| issn | 2399-3642 |
| language | English |
| last_indexed | 2024-03-09T05:30:31Z |
| publishDate | 2023-11-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj.art-a5791267b9534817a20ef741b46632622023-12-03T12:33:13ZengNature PortfolioCommunications Biology2399-36422023-11-016111510.1038/s42003-023-05603-6Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutationsShushu Wei0Niloofar Gopal Pour1Sriram Tiruvadi-Krishnan2Arka Prabha Ray3Naveen Thakur4Matthew T. Eddy5Rajan Lamichhane6Department of Biochemistry & Cellular and Molecular Biology, College of Arts and Sciences, University of TennesseeDepartment of Chemistry, College of Liberal Arts and Sciences, University of FloridaDepartment of Biochemistry & Cellular and Molecular Biology, College of Arts and Sciences, University of TennesseeDepartment of Chemistry, College of Liberal Arts and Sciences, University of FloridaDepartment of Chemistry, College of Liberal Arts and Sciences, University of FloridaDepartment of Chemistry, College of Liberal Arts and Sciences, University of FloridaDepartment of Biochemistry & Cellular and Molecular Biology, College of Arts and Sciences, University of TennesseeAbstract Mutations that constitutively activate G protein-coupled receptors (GPCRs), known as constitutively activating mutations (CAMs), modify cell signaling and interfere with drugs, resulting in diseases with limited treatment options. We utilize fluorescence imaging at the single-molecule level to visualize the dynamic process of CAM-mediated activation of the human A2A adenosine receptor (A2AAR) in real time. We observe an active-state population for all CAMs without agonist stimulation. Importantly, activating mutations significantly increase the population of an intermediate state crucial for receptor activation, notably distinct from the addition of a partner G protein. Activation kinetics show that while CAMs increase the frequency of transitions to the intermediate state, mutations altering sodium sensitivity increase transitions away from it. These findings indicate changes in GPCR function caused by mutations may be predicted based on whether they favor or disfavor formation of an intermediate state, providing a framework for designing receptors with altered functions or therapies that target intermediate states.https://doi.org/10.1038/s42003-023-05603-6 |
| spellingShingle | Shushu Wei Niloofar Gopal Pour Sriram Tiruvadi-Krishnan Arka Prabha Ray Naveen Thakur Matthew T. Eddy Rajan Lamichhane Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations Communications Biology |
| title | Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations |
| title_full | Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations |
| title_fullStr | Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations |
| title_full_unstemmed | Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations |
| title_short | Single-molecule visualization of human A2A adenosine receptor activation by a G protein and constitutively activating mutations |
| title_sort | single molecule visualization of human a2a adenosine receptor activation by a g protein and constitutively activating mutations |
| url | https://doi.org/10.1038/s42003-023-05603-6 |
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