A TPX2 Proteomimetic Has Enhanced Affinity for Aurora-A Due to Hydrocarbon Stapling of a Helix.
A TPX2 Proteomimetic Has Enhanced Affinity for Aurora-A Due to Hydrocarbon Stapling of a Helix.
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DOI：
10.1021/acschembio.6b00727
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发表时间：
2016-11
影响因子：
4
通讯作者：
Yana K Rennie;P. McIntyre;Tito Akindele;R. Bayliss;A. Jamieson
中科院分区：
文献类型：
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作者：
Yana K Rennie;P. McIntyre;Tito Akindele;R. Bayliss;A. Jamieson
利用atp竞争化合物抑制蛋白激酶是药物开发的重要策略。相比之下，尽管有选择性靶向的潜力，但通过破坏蛋白质-蛋白质相互作用来调节激酶的变构调节尚未被广泛采用。Aurora-A激酶调节有丝分裂进入和有丝分裂纺锤体组装，是抗癌治疗的一个有希望的靶点。微管相关蛋白TPX2通过结合两个位点激活Aurora-A。Aurora-A的识别是由TPX2的前43个残基中的两个基序介导的，它们由一个柔性连接体连接。为了表征这三个结构元件的作用，我们制备了一系列TPX2蛋白质组学，并使用等温滴定量热法研究了它们与Aurora-A的结合亲和力。研究人员开发了一种新的钉接TPX2肽，该肽提高了与Aurora-A的结合亲和力，并模拟了TPX2激活Aurora-A自磷酸化的功能。我们得出结论，TPX2的螺旋区域在与Aurora-A结合时折叠，并且该螺旋的稳定并不影响Aurora-A的激活。该研究表明，使用现代合成方法制备这些蛋白质组学物是可行的，它们的生化评价表明，蛋白质组学物作为研究涉及蛋白质内在无序区域的PPIs的工具化合物的力量。
Inhibition of protein kinases using ATP-competitive compounds is an important strategy in drug discovery. In contrast, the allosteric regulation of kinases through the disruption of protein-protein interactions has not been widely adopted, despite the potential for selective targeting. Aurora-A kinase regulates mitotic entry and mitotic spindle assembly and is a promising target for anticancer therapy. The microtubule-associated protein TPX2 activates Aurora-A through binding to two sites. Aurora-A recognition is mediated by two motifs within the first 43 residues of TPX2, connected by a flexible linker. To characterize the contributions of these three structural elements, we prepared a series of TPX2 proteomimetics and investigated their binding affinity for Aurora-A using isothermal titration calorimetry. A novel stapled TPX2 peptide was developed that has improved binding affinity for Aurora-A and mimics the function of TPX2 in activating Aurora-A's autophosphorylation. We conclude that the helical region of TPX2 folds upon binding Aurora-A, and that stabilization of this helix does not compromise Aurora-A activation. This study demonstrates that the preparation of these proteomimetics using modern synthesis methods is feasible and their biochemical evaluation demonstrates the power of proteomimetics as tool compounds for investigating PPIs involving intrinsically disordered regions of proteins.