All-in-One Luminescent Lanthanide Coordination Polymer Nanoprobe for Facile Detection of Protein Kinase Activity
2022
Liu, Baoxia | Zhang, Yaoyao | Hao, Yuanqiang | Zhu, Xu | Zhang, Yintang | Zhou, Yanli | Tan, Hongliang | Xu, Maotian
This study developed a novel luminescent assay for kinase activity using metal–organic coordination polymer nanoparticles (Tb/ATP-Zn) as the probe. Tb/ATP-Zn, self-assembled by adenosine triphosphate (ATP), Zn²⁺, and Tb³⁺, is non-luminescent. Protein kinase A (PKA) can catalyze the transformation of ATP within Tb/ATP-Zn nanoparticles to adenosine diphosphate (ADP), which in turn effectively sensitizes the luminescence of Tb³⁺. Based on this mechanism, Tb/ATP-Zn can realize the facile luminescent “turn-on” sensing of protein kinase activity without the use of external ATP and substrate peptide. Under optimized conditions, the fluorescence intensities of Tb/ATP-Zn at 550 nm are linear with the PKA activity within a range of 0.3–1.5 U·μL–¹. The LOD (S/N = 3) of this method is down to 0.001 U·μL–¹. The presented assay also features high selectivity, long-term stability, fast response, and convenient operation. Furthermore, Tb/ATP-Zn was successfully employed for monitoring PKA activity in cell lysis solutions. Probe Tb/ATP-Zn is thus expectable to be a powerful tool for the practical study of PKA in relevant biological events.
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Эту запись предоставил National Agricultural Library