Small Molecule Probes That Perturb A Protein-protein Interface In Antithrombin.
Small Molecule Probes That Perturb A Protein-protein Interface In Antithrombin.
复制标题
干扰抗凝血酶中蛋白质-蛋白质界面的小分子探针。
DOI：
10.1039/c4sc01295j
复制
发表时间：
2014
期刊：
影响因子：
8.4
通讯作者：
Burgess,Kevin
中科院分区：
文献类型：
--
作者：
Xin,Dongyue;Holzenburg,Andreas;Burgess,Kevin
体外干扰蛋白-蛋白相互作用（PPIs）的小分子探针如果能引起靶蛋白的三级和四级结构发生生物医学上相关的变化，则是有用的。探索关键方向（EKO）策略的应用[j]。化学。Soc。（生物工程学报，2013,135,167-173）表明特异性衍生物可以干扰α-抗凝血酶二聚体中的蛋白-蛋白界面；我们准备并测试了1的特定导数。在这种情况下，它们中的大多数显著加速了单体α-抗凝血酶的寡聚，而α-抗凝血酶在单体状态下是亚稳的。这一论断得到了凝胶电泳（非变性PAGE）和探针诱导的凝血酶催化反应中α-抗凝血酶抑制剂活性丧失的数据的支持。研究了靶化合物诱导α-抗凝血酶寡聚的动力学。结果表明，具有o -苯基保护丝氨酸侧链的探针是该系列催化剂中活性最高的，并通过模拟实验对其原因进行了分析。总的来说，这项研究揭示了第一个小分子的例子之一，这些小分子被设计用于与丝氨酸蛋白（即α-抗凝血酶）寡聚相关的蛋白质-蛋白质界面。这与低聚蛇形蛋白原纤维形成的相关性，与疾病状态称为“蛇形蛋白病”，进行了讨论。
Small molecule probes for perturbing protein–protein interactions (PPIs) in vitro can be useful if they cause the target proteins to undergo biomedically relevant changes to their tertiary and quaternary structures. Application of the Exploring Key Orientations (EKO) strategy (J. Am. Chem. Soc., 2013, 135, 167–173) to a piperidinone–piperidine chemotype 1 indicated specific derivatives were candidates to perturb a protein–protein interface in the α-antithrombin dimer; those particular derivatives of 1 were prepared and tested. In the event, most of them significantly accelerated oligomerization of monomeric α-antithrombin, which is metastable in its monomeric state. This assertion is supported by data from gel electrophoresis (non-denaturing PAGE; throughout) and probe-induced loss of α-antithrombin's inhibitor activity in a reaction catalyzed by thrombin. Kinetics of α-antithrombin oligomerization induced by the target compounds were examined. It was found that probes with O-benzyl-protected serine side-chains are the most active catalysts in the series, and reasons for this, based on modeling experiments, are proposed. Overall, this study reveals one of the first examples of small molecules designed to act at a protein–protein interface relevant to oligomerization of a serpin (i.e. α-antithrombin). The relevance of this to formation of oligomeric serpin fibrils, associated with the disease states known as “serpinopathies”, is discussed.