Direct SUMOylation of M1 muscarinic acetylcholine receptor increases its ligand-binding affinity and signal transduction
| Autor: | Hao Wang, Wei Zhou, Hui Li, Pan-Pan Tan, Juan Li, Liang Zhu, Hongzhuan Chen, Jianrong Xu, Yong-Yao Cui, Yu Qiu, Xuan Zhang |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Carbachol Immunoprecipitation Inositol Phosphates SUMO-1 Protein SUMO protein CHO Cells Molecular Dynamics Simulation Ligands Biochemistry 03 medical and health sciences 0302 clinical medicine Cricetulus Muscarinic acetylcholine receptor Genetics medicine Animals Calcium Signaling Receptor Molecular Biology G protein-coupled receptor Binding Sites Chemistry Receptor Muscarinic M1 Colocalization Sumoylation Cell biology Kinetics 030104 developmental biology Amino Acid Substitution Mutagenesis Site-Directed Signal transduction 030217 neurology & neurosurgery Biotechnology medicine.drug Signal Transduction |
| Zdroj: | FASEB journal : official publication of the Federation of American Societies for Experimental Biology. 33(3) |
| ISSN: | 1530-6860 |
| Popis: | SUMOylation is a significant post-translational modification (PTM) by the small ubiquitin-related modifier (SUMO). Increasing evidence shows SUMOylation regulates GPCR signaling; however, very few GPCRs have been shown to be SUMOylation targets to date. In this study, we identified M1 muscarinic acetylcholine receptor (M1 mAChR), a member of the GPCRs, as a new SUMO substrate. When the mAChR was activated by the agonist carbachol, the colocalization of the M1 mAChR and SUMO-1 protein markedly decreased in immunoprecipitation and immunofluorescence assays. SUMOylation of the M1 mAChR played an important role in increasing the ligand-binding affinity to M1 mAChR, signaling efficiencies, and receptor endocytosis. Through the site-directed mutagenesis approach, K327 was identified as the SUMOylation site of the M1 mAChR. Mutation of the consensus SUMOylation site of the M1 mAChR reduces not only the colocalization of SUMO-1, but also the ligand-binding affinity and signal transduction. The function of M1 mAChR was regulated by SUMOylation through the stabilization of active-state conformation revealed by molecular dynamics simulations. Our results provide evidence that M1 SUMOylation is an important PTM involved in regulation of the affinity for agonists and for activation of signaling pathways.-Xu, J., Tan, P., Li, H., Cui, Y., Qiu, Y., Wang, H., Zhang, X., Li, J., Zhu, L., Zhou, W., Chen, H. Direct SUMOylation of M1 muscarinic acetylcholine receptor increases its ligand-binding affinity and signal transduction. |
| Databáze: | OpenAIRE |
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