Institute of Biochemistry and Center for Molecular Biosciences, University of Innsbruck, Innsbruck, Austria.
IUBMB Life. 2020 Jun;72(6):1168-1174. doi: 10.1002/iub.2241. Epub 2020 Feb 6.
The spectrum of kinase alterations displays distinct functional characteristics and requires kinase mutation-oriented strategies for therapeutic interference. Besides phosphotransferase activity, protein abundance, and intermolecular interactions, particular patient-mutations promote pathological kinase conformations. Despite major advances in identifying lead molecules targeting clinically relevant oncokinase functions, still many kinases are neglected and not part of drug discovery efforts. One explanation is attributed to challenges in tracking kinase activities. Chemical probes are needed to functionally annotate kinase functions, whose activities may not always depend on catalyzing phospho-transfer. Such non-catalytic kinase functions are related to transitions of full-length kinase conformations. Recent findings underline that cell-based reporter systems can be adapted to record conformation changes of kinases. Here, we discuss the possible applications of an extendable kinase conformation (KinCon) reporter toolbox for live-cell recording of kinase states. KinCon is a genetically encoded bioluminescence-based biosensor platform, which can be subjected for measurements of conformation dynamics of mutated kinases upon small molecule inhibitor exposure. We hypothesize that such biosensors can be utilized to delineate the molecular modus operandi for kinase and pseudokinase regulation. This should pave the path for full-length kinase-targeted drug discovery efforts aiming to identify single and combinatory kinase inhibitor therapies with increased specificity and efficacy.
激酶改变的谱显示出独特的功能特征，需要针对激酶突变的策略进行治疗干预。除了磷酸转移酶活性、蛋白质丰度和分子间相互作用外，特定的患者突变会促进病理性激酶构象。尽管在确定针对临床相关致癌激酶功能的先导分子方面取得了重大进展，但仍有许多激酶被忽视，未被纳入药物发现工作中。一种解释归因于跟踪激酶活性的挑战。需要化学探针来对激酶功能进行功能注释，其活性并不总是依赖于催化磷酸转移。这种非催化激酶功能与全长激酶构象的转变有关。最近的发现强调，基于细胞的报告系统可以被改编来记录激酶构象的变化。在这里，我们讨论了可扩展激酶构象（KinCon）报告工具包在活细胞记录激酶状态中的可能应用。KinCon 是一种基于遗传编码的生物发光生物传感器平台，可用于测量小分子抑制剂暴露后突变激酶的构象动力学。我们假设，这种生物传感器可用于描绘激酶和拟激酶调节的分子作用模式。这将为全长激酶靶向药物发现工作铺平道路，旨在确定具有更高特异性和疗效的单一和组合激酶抑制剂治疗方法。
