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通过 ATP 到 ADP 的转换,利用可编程镧系金属-有机骨架增强激酶活性检测。

Enhancing Kinase Activity Detection with a Programmable Lanthanide Metal-Organic Framework via ATP-to-ADP Conversion.

机构信息

Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University, School of Pharmaceutical Sciences, Wuhan University, Wuhan, Hubei 430071, China.

Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), School of Pharmaceutical Sciences, Wuhan University, Wuhan, Hubei 430071, China.

出版信息

Anal Chem. 2024 Jul 23;96(29):12139-12146. doi: 10.1021/acs.analchem.4c02237. Epub 2024 Jul 11.

Abstract

Precise modulation of host-guest interactions between programmable Ln-MOFs (lanthanide metal-organic frameworks) and phosphate analytes holds immense promise for enabling novel functionalities in biosensing. However, the intricate relationship between these functionalities and structures remains largely elusive. Understanding this correlation is crucial for advancing the rational design of fluorescent biosensor technology. Presently, there exists a large research gap concerning the utilization of Ln-MOFsto monitor the conversion of ATP to ADP, which poses a limitation for kinase detection. In this work, we delve into the potential of Ln-MOFs to amplify the fluorescence response during the kinase-mediated ATP-to-ADP conversion. Six Eu-MOFs were synthesized and Eu-TPTC ([1,1':4',1″]-terphenyl-3,3'',5,5''-tetracarboxylic acid) was selected as a ratiometric fluorescent probe, which is most suitable for high-precision detection of creatine kinase activity through the differential response from ATP to ADP. The molecular -level mechanism was confirmed by density functional theory. Furthermore, a simple paper chip-based platform was constructed to realize the fast (20 min) and sensitive (limit of detection is 0.34 U/L) creatine kinase activity detection in biological samples. Ln-MOF-phosphate interactions offer promising avenues for kinase activity assays and hold the potential for precise customization of analytical chemistry.

摘要

精确调控可编程 Ln-MOFs(镧系金属有机骨架)与磷酸盐分析物之间的主客体相互作用,有望为生物传感中的新型功能提供支持。然而,这些功能与结构之间的复杂关系在很大程度上仍难以捉摸。理解这种相关性对于推进荧光生物传感器技术的合理设计至关重要。目前,利用 Ln-MOFs 来监测 ATP 向 ADP 的转化以用于激酶检测方面的研究还存在很大的空白。在这项工作中,我们深入研究了 Ln-MOFs 在激酶介导的 ATP 向 ADP 转化过程中放大荧光响应的潜力。合成了六种 Eu-MOFs,并选择 Eu-TPTC([1,1':4',1″]-三联苯-3,3'',5,5''-四羧酸)作为比率荧光探针,它通过 ATP 到 ADP 的差异响应,最适合用于高精准度地检测肌酸激酶活性。通过密度泛函理论证实了分子水平的机制。此外,构建了一个简单的基于纸芯片的平台,以实现生物样品中肌酸激酶活性的快速(20 分钟)和灵敏(检测限为 0.34 U/L)检测。Ln-MOF-磷酸盐相互作用为激酶活性测定提供了有前景的途径,并有可能精确定制分析化学。

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