Silver Nanoclusters-Based Fluorescence Assay of Protein Kinase Activity and Inhibition

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• 作者：Congcong Shen ; Xiaodong Xia ; Shengqiang Hu ; Minghui Yang ; Jianxiu Wang
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：January 6, 2015
• 年：2015
• 卷：87
• 期：1
• 页码：693-698
• 全文大小：367K
• ISSN：1520-6882

文摘

A simple and sensitive fluorescence method for monitoring the activity and inhibition of protein kinase (PKA) has been developed using polycytosine oligonucleotide (dC₁₂)-templated silver nanoclusters (Ag NCs). Adenosine-5鈥?triphosphate (ATP) was found to enhance the fluorescence of Ag NCs, while the hydrolysis of ATP to adenosine diphosphate (ADP) by PKA decreased the fluorescence of Ag NCs. Compared to the existing methods for kinase activity assay, the developed method does not involve phosphorylation of the substrate peptides, which significantly simplifies the detection procedures. The method exhibits high sensitivity, good selectivity, and wide linear range toward PKA detection. The inhibition effect of kinase inhibitor H-89 on the activity of PKA was also studied. The sensing protocol was also applied to the assay of drug-stimulated activation of PKA in HeLa cell lysates.