Cooperativity between the orthosteric and allosteric ligand binding sites of RORγt
| العنوان: | Cooperativity between the orthosteric and allosteric ligand binding sites of RORγt |
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| المؤلفون: | Rens M J M de Vries, Femke A. Meijer, Luc Brunsveld, Richard G. Doveston, Iris A. Leijten-van de Gevel |
| المساهمون: | Chemical Biology, ICMS Core |
| المصدر: | Proceedings of the National Academy of Sciences of the United States of America (PNAS), 118(6):e2021287118. National Academy of Sciences Proceedings of the National Academy of Sciences of the United States of America |
| بيانات النشر: | National Academy of Sciences, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | genetic structures, Protein Conformation, Allosteric regulation, Molecular Conformation, nuclear receptors, Cooperativity, Molecular Dynamics Simulation, Crystallography, X-Ray, Ligands, Biochemistry, Biophysical Phenomena, drug discovery, 03 medical and health sciences, 0302 clinical medicine, Allosteric Regulation, Humans, 030304 developmental biology, 0303 health sciences, Multidisciplinary, Binding Sites, Drug discovery, Chemistry, structure elucidation, Cooperative binding, ROR gamma t, Nuclear Receptor Subfamily 1, Group F, Member 3, Biological Sciences, Ligand (biochemistry), Förster resonance energy transfer, Nuclear receptor, 030220 oncology & carcinogenesis, Helix, Physical Sciences, Biophysics, RORγt, allosteric modulators, Allosteric Site, Protein Binding |
| الوصف: | Significance RORγt is a nuclear receptor associated with several diseases. Various synthetic ligands have been developed that target the canonical orthosteric or a second, allosteric pocket of RORγt. We show that orthosteric and allosteric ligands can simultaneously bind to RORγt and that their potency is positively influenced by the other ligand, a phenomenon called cooperative dual ligand binding. The mechanism behind cooperative binding in proteins is poorly understood, primarily due to the lack of structural data. We solved 12 crystal structures of RORγt, simultaneously bound to various orthosteric and allosteric ligands. In combination with molecular dynamics, we reveal a mechanism responsible for the cooperative binding behavior. Our comprehensive structural studies provide unique insights into how cooperative binding occurs in proteins. Cooperative ligand binding is an important phenomenon in biological systems where ligand binding influences the binding of another ligand at an alternative site of the protein via an intramolecular network of interactions. The underlying mechanisms behind cooperative binding remain poorly understood, primarily due to the lack of structural data of these ternary complexes. Using time-resolved fluorescence resonance energy transfer (TR-FRET) studies, we show that cooperative ligand binding occurs for RORγt, a nuclear receptor associated with the pathogenesis of autoimmune diseases. To provide the crucial structural insights, we solved 12 crystal structures of RORγt simultaneously bound to various orthosteric and allosteric ligands. The presence of the orthosteric ligand induces a clamping motion of the allosteric pocket via helices 4 to 5. Additional molecular dynamics simulations revealed the unusual mechanism behind this clamping motion, with Ala355 shifting between helix 4 and 5. The orthosteric RORγt agonists regulate the conformation of Ala355, thereby stabilizing the conformation of the allosteric pocket and cooperatively enhancing the affinity of the allosteric inverse agonists. |
| اللغة: | English |
| تدمد: | 1091-6490 0027-8424 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4af3aeb1394211469396b3c50ee1046cTest https://doi.org/10.1073/pnas.2021287118Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....4af3aeb1394211469396b3c50ee1046c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 00278424 |
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