<br>ATP 竞争性和新型非竞争性 Abbapolin 抑制剂诱导的 Polo 样激酶 1 构象和细胞内稳定性变化的结构基础,ACS Chemical Biology

当前位置： X-MOL 学术 › ACS Chem. Biol. › 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.)

ATP 竞争性和新型非竞争性 Abbapolin 抑制剂诱导的 Polo 样激酶 1 构象和细胞内稳定性变化的结构基础
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2023-07-11 , DOI: 10.1021/acschembio.3c00269
Danda Chapagai ₁ , George Merhej ₁ , Campbell McInnes ₁ , Michael D Wyatt ₁

Affiliation

Polo 样激酶 1 (PLK1) 是一种必需蛋白激酶，在有丝分裂进展中具有多种作用。 PLK1 由激酶结构域 (KD) 和磷酸肽结合 polobox 结构域 (PBD) 组成，负责底物识别和亚细胞定位。 PLK1 的调节涉及 KD 和 PBD 相互作用的自抑制构象。我们之前的工作发现了称为 abbapolins 的 PBD 结合分子，可抑制 PLK1 底物的细胞磷酸化并诱导细胞内 PLK1 的丢失。在这里，我们描述了 abbapolin 活性与 KD 抑制剂的活性比较，以深入了解 PLK1 的构象特征。通过细胞热位移测定测量，abbabalins 产生配体诱导的 PLK1 热稳定性。相反，KD 抑制剂降低了可溶性 PLK1，这表明催化位点结合导致 PLK1 构象热稳定性较差。全长 PLK1 和 KD 抑制剂的结合测量也证明了构象变化。有趣的是，KD 与 PBD 结合的细胞后果形成鲜明对比，因为 KD 结合会导致细胞内 PLK1 的积累，而 PBD 结合会导致核 PLK1 的显着损失。这些数据与 KD 结合剂缓解 PLK1 自身抑制作用一致；使用 AlphaFold 预测的催化结构域和全长 PLK1 的结构对这些观察结果进行了解释。总的来说，这些结果凸显了靶向 PLK1 的一个未被充分认识的方面，即 KD 与 PBD 结合引起的构象扰动。除了对 PBD 结合配体具有重要意义外，这些观察结果还对 ATP 竞争性 PLK1 抑制剂的开发具有影响，因为催化抑制剂可能反过来促进 PLK1 非催化功能，这可能解释了它们迄今为止缺乏临床疗效的原因。

"点击查看英文标题和摘要"

Structural Basis for Variations in Polo-like Kinase 1 Conformation and Intracellular Stability Induced by ATP-Competitive and Novel Noncompetitive Abbapolin Inhibitors

Polo-like kinase 1 (PLK1) is an essential protein kinase with multiple roles in mitotic progression. PLK1 consists of a kinase domain (KD) and a phosphopeptide-binding polobox domain (PBD), which is responsible for substrate recognition and subcellular localization. The regulation of PLK1 involves an autoinhibitory conformation in which KD and PBD interact. Our previous work identified PBD-binding molecules termed abbapolins that inhibit the cellular phosphorylation of a PLK1 substrate and induce the loss of intracellular PLK1. Here, we describe a comparison of the abbapolin activity with that of KD inhibitors to gain insight into conformational features of PLK1. As measured by a cellular thermal shift assay, abbapolins produce ligand-induced thermal stabilization of PLK1. In contrast, KD inhibitors decreased the soluble PLK1, suggesting that catalytic-site binding causes a less thermally stable PLK1 conformation. Binding measurements with full-length PLK1 and a KD inhibitor also demonstrated a conformational change. Interestingly, the cellular consequences of KD versus PBD engagement contrast as KD binding causes the accumulation of intracellular PLK1, whereas PBD binding produces a striking loss of nuclear PLK1. These data are consistent with the relief of autoinhibited PLK1 by KD binders; an explanation for these observations is presented using structures for the catalytic domain and full-length PLK1 predicted by AlphaFold. Collectively, the results highlight an underappreciated aspect of targeting PLK1, namely, conformational perturbations induced by KD versus PBD binding. In addition to their significance for PBD-binding ligands, these observations have implications for the development of ATP-competitive PLK1 inhibitors because catalytic inhibitors may conversely promote PLK1 noncatalytic functions, which may explain their lack of clinical efficacy to date.

更新日期：2023-07-11

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南