Kugler Valentina, Schwaighofer Selina, Feichtner Andreas, Enzler Florian, Fleischmann Jakob, Strich Sophie, Schwarz Sarah, Wilson Rebecca, Tschaikner Philipp, Troppmair Jakob, Sexl Veronika, Meier Pascal, Kaserer Teresa, Stefan Eduard
Institute for Molecular Biology and Center for Molecular Biosciences Innsbruck (CMBI), University of Innsbruck, Innsbruck, Austria.
Tyrolean Cancer Research Institute (TKFI), Innsbruck, Austria.
Elife. 2024 Aug 1;13:RP94755. doi: 10.7554/eLife.94755.
Protein kinases act as central molecular switches in the control of cellular functions. Alterations in the regulation and function of protein kinases may provoke diseases including cancer. In this study we investigate the conformational states of such disease-associated kinases using the high sensitivity of the kinase conformation (KinCon) reporter system. We first track BRAF kinase activity conformational changes upon melanoma drug binding. Second, we also use the KinCon reporter technology to examine the impact of regulatory protein interactions on LKB1 kinase tumor suppressor functions. Third, we explore the conformational dynamics of RIP kinases in response to TNF pathway activation and small molecule interactions. Finally, we show that CDK4/6 interactions with regulatory proteins alter conformations which remain unaffected in the presence of clinically applied inhibitors. Apart from its predictive value, the KinCon technology helps to identify cellular factors that impact drug efficacies. The understanding of the structural dynamics of full-length protein kinases when interacting with small molecule inhibitors or regulatory proteins is crucial for designing more effective therapeutic strategies.
蛋白激酶在细胞功能控制中充当核心分子开关。蛋白激酶调控和功能的改变可能引发包括癌症在内的疾病。在本研究中，我们利用激酶构象（KinCon）报告系统的高灵敏度来研究此类与疾病相关激酶的构象状态。我们首先追踪黑色素瘤药物结合后BRAF激酶活性的构象变化。其次，我们还使用KinCon报告技术来研究调节蛋白相互作用对LKB1激酶肿瘤抑制功能的影响。第三，我们探究RIP激酶响应TNF途径激活和小分子相互作用时的构象动力学。最后，我们表明CDK4/6与调节蛋白的相互作用会改变构象，而在临床应用抑制剂存在的情况下这些构象保持不变。除了其预测价值外，KinCon技术有助于识别影响药物疗效的细胞因子。了解全长蛋白激酶与小分子抑制剂或调节蛋白相互作用时的结构动力学对于设计更有效的治疗策略至关重要。
