Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor
Biased agonism, the ability of agonists to differentially activate downstream signaling pathways by stabilizing specific receptor conformations, is a key issue for G protein-coupled receptor (GPCR) signaling. The C-terminal domain might influence this functional selectivity of GPCRs as it engages G...
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| Format: | Article |
| Language: | English |
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Elsevier
2020-01-01
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| Series: | Computational and Structural Biotechnology Journal |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2001037020304128 |
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| author | Gemma Navarro Angel Gonzalez Stefano Campanacci Rafael Rivas-Santisteban Irene Reyes-Resina Nil Casajuana-Martin Arnau Cordomí Leonardo Pardo Rafael Franco |
| author_facet | Gemma Navarro Angel Gonzalez Stefano Campanacci Rafael Rivas-Santisteban Irene Reyes-Resina Nil Casajuana-Martin Arnau Cordomí Leonardo Pardo Rafael Franco |
| author_sort | Gemma Navarro |
| collection | DOAJ |
| description | Biased agonism, the ability of agonists to differentially activate downstream signaling pathways by stabilizing specific receptor conformations, is a key issue for G protein-coupled receptor (GPCR) signaling. The C-terminal domain might influence this functional selectivity of GPCRs as it engages G proteins, GPCR kinases, β-arrestins, and several other proteins. Thus, the aim of this paper is to compare the agonist-dependent selectivity for intracellular pathways in a heterologous system expressing the full-length (A2AR) and a C-tail truncated (A2AΔ40R lacking the last 40 amino acids) adenosine A2A receptor, a GPCR that is already targeted in Parkinson’s disease using a first-in-class drug. Experimental data such as ligand binding, cAMP production, β-arrestin recruitment, ERK1/2 phosphorylation and dynamic mass redistribution assays, which correspond to different aspects of signal transduction, were measured upon the action of structurally diverse compounds (the agonists adenosine, NECA, CGS-21680, PSB-0777 and LUF-5834 and the SCH-58261 antagonist) in cells expressing A2AR and A2AΔ40R. The results show that taking cAMP levels and the endogenous adenosine agonist as references, the main difference in bias was obtained with PSB-0777 and LUF-5834. The C-terminus is dispensable for both G-protein and β-arrestin recruitment and also for MAPK activation. Unrestrained molecular dynamics simulations, at the μs timescale, were used to understand the structural arrangements of the binding cavity, triggered by these chemically different agonists, facilitating G protein binding with different efficacy. |
| first_indexed | 2024-12-16T10:18:41Z |
| format | Article |
| id | doaj.art-b15d52c3368248e7b461d60c36c91d11 |
| institution | Directory Open Access Journal |
| issn | 2001-0370 |
| language | English |
| last_indexed | 2024-12-16T10:18:41Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Computational and Structural Biotechnology Journal |
| spelling | doaj.art-b15d52c3368248e7b461d60c36c91d112022-12-21T22:35:22ZengElsevierComputational and Structural Biotechnology Journal2001-03702020-01-011827232732Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptorGemma Navarro0Angel Gonzalez1Stefano Campanacci2Rafael Rivas-Santisteban3Irene Reyes-Resina4Nil Casajuana-Martin5Arnau Cordomí6Leonardo Pardo7Rafael Franco8Dept. Biochemistry and Physiology, Faculty of Pharmacy and Food Science. Universitat de Barcelona. Barcelona, Spain; Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas. Instituto de Salud Carlos III, Madrid, SpainLaboratori de Medicina Computacional, Unitat de Bioestadística, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain d Faculty of Chemistry, Universitat de Barcelona, Barcelona, SpainDept. Biochemistry and Molecular Biomedicine. School of Biology. Universitat de Barcelona. Barcelona. SpainDept. Biochemistry and Physiology, Faculty of Pharmacy and Food Science. Universitat de Barcelona. Barcelona, Spain; Dept. Biochemistry and Molecular Biomedicine. School of Biology. Universitat de Barcelona. Barcelona. SpainCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas. Instituto de Salud Carlos III, Madrid, Spain; Dept. Biochemistry and Molecular Biomedicine. School of Biology. Universitat de Barcelona. Barcelona. SpainLaboratori de Medicina Computacional, Unitat de Bioestadística, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain d Faculty of Chemistry, Universitat de Barcelona, Barcelona, SpainLaboratori de Medicina Computacional, Unitat de Bioestadística, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain d Faculty of Chemistry, Universitat de Barcelona, Barcelona, SpainLaboratori de Medicina Computacional, Unitat de Bioestadística, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain d Faculty of Chemistry, Universitat de Barcelona, Barcelona, SpainCentro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas. Instituto de Salud Carlos III, Madrid, Spain; School of Chemistry. Universitat de Barcelona. Barcelona. Spain; Corresponding author at: School of Chemistry, University of Barcelona, Diagonal 643, 08028 Barcelona, Spain.Biased agonism, the ability of agonists to differentially activate downstream signaling pathways by stabilizing specific receptor conformations, is a key issue for G protein-coupled receptor (GPCR) signaling. The C-terminal domain might influence this functional selectivity of GPCRs as it engages G proteins, GPCR kinases, β-arrestins, and several other proteins. Thus, the aim of this paper is to compare the agonist-dependent selectivity for intracellular pathways in a heterologous system expressing the full-length (A2AR) and a C-tail truncated (A2AΔ40R lacking the last 40 amino acids) adenosine A2A receptor, a GPCR that is already targeted in Parkinson’s disease using a first-in-class drug. Experimental data such as ligand binding, cAMP production, β-arrestin recruitment, ERK1/2 phosphorylation and dynamic mass redistribution assays, which correspond to different aspects of signal transduction, were measured upon the action of structurally diverse compounds (the agonists adenosine, NECA, CGS-21680, PSB-0777 and LUF-5834 and the SCH-58261 antagonist) in cells expressing A2AR and A2AΔ40R. The results show that taking cAMP levels and the endogenous adenosine agonist as references, the main difference in bias was obtained with PSB-0777 and LUF-5834. The C-terminus is dispensable for both G-protein and β-arrestin recruitment and also for MAPK activation. Unrestrained molecular dynamics simulations, at the μs timescale, were used to understand the structural arrangements of the binding cavity, triggered by these chemically different agonists, facilitating G protein binding with different efficacy.http://www.sciencedirect.com/science/article/pii/S2001037020304128G protein coupled receptorsAdenosine A2A receptorFunctional selectivityG protein bindingβ-Arrestin recruitmentMolecular dynamic simulations |
| spellingShingle | Gemma Navarro Angel Gonzalez Stefano Campanacci Rafael Rivas-Santisteban Irene Reyes-Resina Nil Casajuana-Martin Arnau Cordomí Leonardo Pardo Rafael Franco Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor Computational and Structural Biotechnology Journal G protein coupled receptors Adenosine A2A receptor Functional selectivity G protein binding β-Arrestin recruitment Molecular dynamic simulations |
| title | Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor |
| title_full | Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor |
| title_fullStr | Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor |
| title_full_unstemmed | Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor |
| title_short | Experimental and computational analysis of biased agonism on full-length and a C-terminally truncated adenosine A2A receptor |
| title_sort | experimental and computational analysis of biased agonism on full length and a c terminally truncated adenosine a2a receptor |
| topic | G protein coupled receptors Adenosine A2A receptor Functional selectivity G protein binding β-Arrestin recruitment Molecular dynamic simulations |
| url | http://www.sciencedirect.com/science/article/pii/S2001037020304128 |
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