Key Laboratory of Public Health Safety of Hebei Province, School of Public Health, Hebei University, Baoding, 071002, PR China.
Key Laboratory of Public Health Safety of Hebei Province, School of Public Health, Hebei University, Baoding, 071002, PR China; Key Laboratory of Medicinal Chemistry and Molecular Diagnosis of Ministry of Education, College of Pharmaceutical Sciences, Hebei University, Baoding, 071002, PR China.
Biosens Bioelectron. 2022 May 1;203:114055. doi: 10.1016/j.bios.2022.114055. Epub 2022 Feb 1.
Protein kinases play crucial regulatory roles in the physiological activities in the human body. Understanding protein kinase activity and its inhibition is essential for the management of human diseases. Considering the limitations of the existing protein kinase-related analysis methods, the aim of the present study was to develop a fluorescent biosensor based on Eu(BTC) (HO) (BTC = 1,3,5-Benzenetricarboxylic acid) for evaluating protein kinase activity and the relevant inhibitors. A fluorophore-labelled substrate polypeptide was phosphorylated under the catalysis of protein kinase. This phosphorylated peptide can be coordinated explicitly with the europium site of Eu(BTC) (HO) to detect the protein kinase. The developed biosensor performed well, with a detection limit of 0.00003 U μL, and it showed good selectivity and universality. Protein kinase activity could also be detected in MCF-7 cells using this method. Furthermore, in terms of inhibitor screening using the Eu(BTC) (HO)-based sensor, both H-89 and ellagic acid were found to inhibit protein kinase activity with IC values of 1.09 and 19.88 nmol L, respectively. Overall, this biosensor has broad application prospects in monitoring and controlling protein kinase activity.
蛋白激酶在人体的生理活动中发挥着至关重要的调节作用。了解蛋白激酶的活性及其抑制作用对于人类疾病的治疗至关重要。鉴于现有蛋白激酶相关分析方法的局限性，本研究旨在开发一种基于 Eu(BTC)(HO)(BTC=1,3,5-苯三甲酸)的荧光生物传感器，用于评估蛋白激酶活性及其相关抑制剂。在蛋白激酶的催化作用下，荧光标记的底物多肽被磷酸化。这种磷酸化肽可以与 Eu(BTC)(HO)的 Eu 位点明确配位，从而检测蛋白激酶。所开发的生物传感器性能良好，检测限为 0.00003 U μL，具有良好的选择性和通用性。该方法还可用于 MCF-7 细胞中的蛋白激酶活性检测。此外，使用基于 Eu(BTC)(HO)的传感器进行抑制剂筛选时，发现 H-89 和鞣花酸对蛋白激酶活性的抑制作用，IC 值分别为 1.09 和 19.88 nmol L。总的来说，这种生物传感器在监测和控制蛋白激酶活性方面具有广阔的应用前景。
