The Substrate-Activity-Screening methodology applied to receptor tyrosine kinases: A proof-of-concept study
The Substrate-Activity-Screening methodology applied to receptor tyrosine kinases: A proof-of-concept study
复制标题
DOI：
10.1016/j.ejmech.2012.08.038
复制
发表时间：
2012-11-01
影响因子：
6.7
通讯作者：
Seebach, Dieter
中科院分区：
文献类型：
--
作者：
Chapelat, Julien;Berst, Frederic;Seebach, Dieter
蛋白激酶因其参与信号转导通路而被广泛认为是重要的治疗靶点。这些途径受到严格的控制和调节，特别是通过激酶选择性磷酸化一组确定的底物的能力。由于激酶介导的磷酸化异常，各种各样的疾病都可能出现，许多激酶抑制剂已经成为现代药典的关键成分。虽然“传统的”激酶抑制剂通常通过阻止激酶和ATP之间的相互作用来起作用，从而阻止底物磷酸化，但另一种方法是破坏激酶与其下游伙伴之间的蛋白质相互作用。为了便于确定底物位点抑制方法的潜在化学起点，我们希望研究底物活性筛选在激酶中的应用。我们在此报告一项概念验证研究，在一个模型酪氨酸激酶上，该方法的关键要求可以得到满足。也就是说，使用多肽作为模型底物，我们表明可以使用简单的adp积累测定来监测底物效率，并且可以以模块化的方式优化效率。更重要的是，我们证明了结构效率关系在底物转化为抑制剂时转化为结构活性关系。(C) 2012 Elsevier Masson SAS。版权所有。
Protein kinases are widely recognized as important therapeutic targets due to their involvement in signal transduction pathways. These pathways are tightly controlled and regulated, notably by the ability of kinases to selectively phosphorylate a defined set of substrates. A wide variety of disorders can arise as a consequence of abnormal kinase-mediated phosphorylation and numerous kinase inhibitors have earned their place as key components of the modern pharmacopeia. Although "traditional" kinase inhibitors typically act by preventing the interaction between the kinase and ATP, thus stopping substrate phosphorylation, an alternative approach consists in disrupting the protein protein interaction between the kinase and its downstream partners. In order to facilitate the identification of potential chemical starting points for substrate-site inhibition approaches, we desired to investigate the application of Substrate Activity Screening to kinases. We herein report a proof-of-concept study demonstrating, on a model tyrosine kinase, that the key requirements of this methodology can be met. Namely, using peptides as model substrates, we show that a simple ADP-accumulation assay can be used to monitor substrate efficiency and that efficiency can be optimized in a modular manner. More importantly, we demonstrate that structure efficiency relationships translate into structure activity relationships upon conversion of the substrates into inhibitors. (C) 2012 Elsevier Masson SAS. All rights reserved.