Cooperativity between the orthosteric and allosteric ligand binding sites of RORγt.
Cooperativity between the orthosteric and allosteric ligand binding sites of RORγt.
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DOI：
10.1073/pnas.2021287118
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发表时间：
2021-02-09
影响因子：
11.1
通讯作者：
Brunsveld L
中科院分区：
文献类型：
--
作者：
de Vries RMJM;Meijer FA;Doveston RG;Leijten-van de Gevel IA;Brunsveld L
RoRγt是一种与多种疾病相关的核受体。针对RORγt的正构体或第二个变构口袋的各种合成配体已经被开发出来。我们发现，正构体和变构配体可以同时与RoRγt结合，并且它们的效力受到另一种配体的正向影响，这种现象被称为合作双配体结合。蛋白质协同结合背后的机制还不是很清楚，主要是因为缺乏结构数据。我们解决了RoRγt的12个晶体结构，同时结合了各种正构和变构配体。结合分子动力学，我们揭示了协同结合行为的机理。我们全面的结构研究为蛋白质中如何发生协同结合提供了独特的见解。协同配基结合是生物系统中的一种重要现象，在生物系统中，配基结合通过分子内相互作用网络影响另一个配基在蛋白质的另一个位置的结合。合作结合背后的潜在机制仍然知之甚少，主要是因为缺乏这些三元络合物的结构数据。利用时间分辨荧光共振能量转移(TRRET)研究，我们证明了与自身免疫性疾病发病相关的核受体RoRγt发生了协同配体结合。为了提供关键的结构见解，我们解决了RoRγt同时与各种正构和变构配体结合的12个晶体结构。正构配体的存在导致变构口袋通过螺旋4到5的夹持运动。额外的分子动力学模拟揭示了这种夹持运动背后的不寻常机制，Ala355在螺旋4和5之间移动。正构RoRγt激动剂调节Ala355的构象，从而稳定变构口袋的构象，协同增强变构反向激动剂的亲和力。
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.
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影响因子：
64.5
作者：
Ivanov, Ivaylo I.;McKenzie, Brent S.;Littman, Dan R.
通讯作者：
Littman, Dan R.
影响因子：
2.7
作者：
Fauber, Benjamin P.;Gobbi, Alberto;Wong, Harvey
通讯作者：
Wong, Harvey
HTRF：一项针对药物发现量身定制的技术 - 对理论方面和最新应用的回顾。
DOI：
10.2174/1875397300903010022
发表时间：
2009-05-28
期刊：
Current chemical genomics
影响因子：
--
作者：
Degorce F;Card A;Soh S;Trinquet E;Knapik GP;Xie B
通讯作者：
Xie B
影响因子：
4.4
作者：
Bussi, Giovanni;Donadio, Davide;Parrinello, Michele
通讯作者：
Parrinello, Michele
影响因子：
2.7
作者：
Cyr, Patrick;Bronner, Sarah M.;Crawford, James J.
通讯作者：
Crawford, James J.