von Glasenapp Victoria, C Almeida Ana, Chang Dalu, Gasic Ivana, Winssinger Nicolas, Gotta Monica
Department of Physiology and Metabolism, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
NCCR Chemical Biology, University of Geneva, Geneva, Switzerland.
Nat Commun. 2025 Feb 19;16(1):1599. doi: 10.1038/s41467-025-56746-5.
The ability to control the activity of kinases spatially and temporally is essential to elucidate the role of signalling pathways in development and physiology. Progress in this direction has been hampered by the lack of tools to manipulate kinase activity in a highly controlled manner in vivo. Here we report a strategy to modify BI2536, the well characterized inhibitor of the conserved and essential mitotic kinase Polo-like kinase 1 (Plk1). We introduce the same coumarin photolabile protecting group (PPG) at two positions of the inhibitor. At one position, the coumarin prevents the interaction with Plk1, at the second it masks an added carboxylic acid, important for cellular retention. Exposure to light results in removal of both PPGs, leading to the activation of the inhibitor and its trapping inside cells. We demonstrate the efficacy of the caged inhibitor in three-dimensional spheroid cultures: by uncaging it with a single light pulse, we can inhibit Plk1 and arrest cell division, a highly dynamic process, with spatio-temporal control. Our design can be applied to other small molecules, providing a solution to control their activity in living cells with unprecedented precision.
在空间和时间上控制激酶活性的能力对于阐明信号通路在发育和生理学中的作用至关重要。由于缺乏在体内以高度可控方式操纵激酶活性的工具，这一方向的进展受到了阻碍。在此，我们报告了一种修饰BI2536的策略，BI2536是保守且必需的有丝分裂激酶Polo样激酶1（Plk1）的特征明确的抑制剂。我们在抑制剂的两个位置引入了相同的香豆素光不稳定保护基团（PPG）。在一个位置，香豆素阻止与Plk1的相互作用，在第二个位置，它掩盖了一个添加的羧酸，这对细胞保留很重要。暴露于光会导致两个PPG都被去除，从而导致抑制剂的激活及其在细胞内的捕获。我们在三维球体培养中证明了笼化抑制剂的功效：通过用单个光脉冲解开笼化，我们可以抑制Plk1并阻止细胞分裂，这是一个高度动态的过程，并具有时空控制。我们的设计可应用于其他小分子，以前所未有的精度提供一种在活细胞中控制其活性的解决方案。
