当前位置：
X-MOL 学术
›
Angew. Chem. Int. Ed.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Unraveling a Ligand-Induced Twist of a Homodimeric Enzyme by Pulsed Electron–Electron Double Resonance
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2021-08-12 , DOI: 10.1002/anie.202108179 Dzung Nguyen 1 , Dinar Abdullin 2 , Caspar A Heubach 2 , Toni Pfaffeneder 3 , Andreas Nguyen 1 , Andreas Heine 1 , Klaus Reuter 1 , François Diederich 3 , Olav Schiemann 2 , Gerhard Klebe 1
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2021-08-12 , DOI: 10.1002/anie.202108179 Dzung Nguyen 1 , Dinar Abdullin 2 , Caspar A Heubach 2 , Toni Pfaffeneder 3 , Andreas Nguyen 1 , Andreas Heine 1 , Klaus Reuter 1 , François Diederich 3 , Olav Schiemann 2 , Gerhard Klebe 1
Affiliation
|
Mechanistic insights into protein–ligand interactions can yield chemical tools for modulating protein function and enable their use for therapeutic purposes. For the homodimeric enzyme tRNA-guanine transglycosylase (TGT), a putative virulence target of shigellosis, ligand binding has been shown by crystallography to transform the functional dimer geometry into an incompetent twisted one. However, crystallographic observation of both end states does neither verify the ligand-induced transformation of one dimer into the other in solution nor does it shed light on the underlying transformation mechanism. We addressed these questions in an approach that combines site-directed spin labeling (SDSL) with distance measurements based on pulsed electron–electron double resonance (PELDOR or DEER) spectroscopy. We observed an equilibrium between the functional and twisted dimer that depends on the type of ligand, with a pyranose-substituted ligand being the most potent one in shifting the equilibrium toward the twisted dimer. Our experiments suggest a dissociation–association mechanism for the formation of the twisted dimer upon ligand binding.
中文翻译：
通过脉冲电子-电子双共振解开配体诱导的同型二聚酶扭曲
对蛋白质-配体相互作用的机制洞察可以产生用于调节蛋白质功能的化学工具，并使其用于治疗目的。对于同源二聚体酶 tRNA-鸟嘌呤转糖基酶 (TGT)，志贺氏菌病的假定毒力目标，晶体学显示配体结合可将功能性二聚体几何结构转化为无能的扭曲几何结构。然而，两种终态的晶体学观察既不能证实配体诱导的溶液中一种二聚体向另一种二聚体的转化，也不能揭示潜在的转化机制。我们通过将定点自旋标记 (SDSL) 与基于脉冲电子-电子双共振（PELDOR 或 DEER）光谱的距离测量相结合的方法解决了这些问题。我们观察到功能性二聚体和扭曲二聚体之间的平衡取决于配体的类型，吡喃糖取代的配体是将平衡向扭曲二聚体转移的最有效的配体。我们的实验表明了在配体结合后形成扭曲二聚体的解离-缔合机制。
更新日期：2021-10-12
中文翻译：
通过脉冲电子-电子双共振解开配体诱导的同型二聚酶扭曲
对蛋白质-配体相互作用的机制洞察可以产生用于调节蛋白质功能的化学工具，并使其用于治疗目的。对于同源二聚体酶 tRNA-鸟嘌呤转糖基酶 (TGT)，志贺氏菌病的假定毒力目标，晶体学显示配体结合可将功能性二聚体几何结构转化为无能的扭曲几何结构。然而，两种终态的晶体学观察既不能证实配体诱导的溶液中一种二聚体向另一种二聚体的转化，也不能揭示潜在的转化机制。我们通过将定点自旋标记 (SDSL) 与基于脉冲电子-电子双共振（PELDOR 或 DEER）光谱的距离测量相结合的方法解决了这些问题。我们观察到功能性二聚体和扭曲二聚体之间的平衡取决于配体的类型，吡喃糖取代的配体是将平衡向扭曲二聚体转移的最有效的配体。我们的实验表明了在配体结合后形成扭曲二聚体的解离-缔合机制。
京公网安备 11010802027423号