ATP-competitive inhibitors of PI3K enzymes demonstrate an isoform selective dual action by controlling membrane binding.

IF 4.4 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemical Journal Pub Date : 2024-11-01 DOI:10.1042/BCJ20240479
Grace Q Gong, Glenn Robert Masson, Woo-Jeong Jeong Lee, James Mj Dickson, Jackie D Kendall, Manoj K Rathinaswamy, Christina M Buchanan, Martin Middleditch, Brady Owen, Julie A Spicer, Gordon W Rewcastle, William A Denny, John E Burke, Peter R Shepherd, Roger L Williams, Jack U Flanagan
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Abstract

PI3Kα, consisting of the p110α isoform of the catalytic subunit of PI 3-kinase (encoded by PIK3CA) and the p85α regulatory subunit (encoded by PI3KR1) is activated by growth factor receptors. The identification of common oncogenic mutations in PIK3CA has driven the development of many inhibitors that bind to the ATP-binding site in the p110α subunit. Upon activation, PI3Kα undergoes conformational changes that promote its membrane interaction and catalytic activity, yet the effects of ATP-site directed inhibitors on the PI3Kα membrane interaction are unknown. Using FRET and Biolayer Interferometry assays, we show that a class of ATP-site directed inhibitors represented by GSK2126458 block the growth factor activated PI3KαWT membrane interaction, an activity dependent on the ligand forming specific ATP-site interactions. The membrane interaction for hot spot oncogenic mutations that bypass normal p85α regulatory mechanisms was insensitive to GSK2126458, while GSK2126458 could regulate mutations found outside of these hot spot regions. Our data show that the effect of GSK126458 on the membrane interaction requires the enzyme to revert from its growth factor activated state to a basal state. We find that an ATP substrate analogue can increase the wild type PI3Kα membrane interaction, uncovering a substrate based regulatory event that can be mimicked by different inhibitor chemotypes. Our findings, together with the discovery of small molecule allosteric activators of PI3Kα illustrate that PI3Kα membrane interactions can be modulated by factors related to ligand binding both within the ATP site and at allosteric sites.

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PI3K 酶的 ATP 竞争性抑制剂通过控制膜结合,显示出同工酶选择性的双重作用。
PI3Kα 由 PI 3-kinase 催化亚基的 p110α 异构体(由 PIK3CA 编码)和 p85α 调节亚基(由 PI3KR1 编码)组成,由生长因子受体激活。PIK3CA 常见致癌突变的发现推动了许多与 p110α 亚基中的 ATP 结合位点结合的抑制剂的开发。激活后,PI3Kα会发生构象变化,从而促进其膜相互作用和催化活性,但ATP位点定向抑制剂对PI3Kα膜相互作用的影响尚不清楚。我们利用 FRET 和生物层干涉测量法检测表明,以 GSK2126458 为代表的一类 ATP 位点定向抑制剂会阻断生长因子激活的 PI3KαWT 膜相互作用,这种活性依赖于配体形成特定的 ATP 位点相互作用。绕过正常 p85α 调节机制的热点致癌突变的膜相互作用对 GSK2126458 不敏感,而 GSK2126458 能调节这些热点区域之外的突变。我们的数据显示,GSK126458 对膜相互作用的影响需要酶从生长因子激活状态恢复到基础状态。我们发现 ATP 底物类似物能增加野生型 PI3Kα 的膜相互作用,从而揭示了一种基于底物的调控事件,不同的抑制剂化学类型可以模拟这种调控事件。我们的发现以及 PI3Kα 的小分子异构激活剂的发现说明,PI3Kα 膜相互作用可受 ATP 位点和异构位点内配体结合相关因素的调节。
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来源期刊
Biochemical Journal
Biochemical Journal 生物-生化与分子生物学
CiteScore
8.00
自引率
0.00%
发文量
255
审稿时长
1 months
期刊介绍: Exploring the molecular mechanisms that underpin key biological processes, the Biochemical Journal is a leading bioscience journal publishing high-impact scientific research papers and reviews on the latest advances and new mechanistic concepts in the fields of biochemistry, cellular biosciences and molecular biology. The Journal and its Editorial Board are committed to publishing work that provides a significant advance to current understanding or mechanistic insights; studies that go beyond observational work using in vitro and/or in vivo approaches are welcomed. Painless publishing: All papers undergo a rigorous peer review process; however, the Editorial Board is committed to ensuring that, if revisions are recommended, extra experiments not necessary to the paper will not be asked for. Areas covered in the journal include: Cell biology Chemical biology Energy processes Gene expression and regulation Mechanisms of disease Metabolism Molecular structure and function Plant biology Signalling
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