Abstract：
T4 polynucleotide kinase (T4 PNK) helps with DNA recombination and repair. In this work, a phosphate pillar[5]arene@palladium nanoparticles@reduced graphene oxide nanocomposite (PP5@PdNPs@rGO)-based electrochemical biosensor was created to identify T4 PNK activities. The PP5 used to complex toluidine blue (TB) guest molecules is water-soluble. With T4 PNK and ATP, the substrate DNA, which included a 5\'-hydroxyl group, initially self-assembled over the gold electrode surface by chemical adsorption of the thiol units. Strong phosphate-Zr4+-phosphate chemistry allowed Zr4+ to act as a bridge between phosphorylated DNA and PP5@PdNPs@rGO. Through a supramolecular host-guest recognition connection, TB molecules were able to penetrate the PP5 cavity, where they produced a stronger electrochemical response. With a 5 × 10-7 U mL-1 detection limit, the electrochemical signal is linear in the 10-6 to 1 U mL-1 T4 PNK concentration range. It was also effective in measuring HeLa cell lysate-related PNK activities and screening PNK inhibitors. Nucleotide kinase-target drug development, clinical diagnostics, and screening for inhibitors all stand to benefit greatly from the suggested technology, which offers a unique sensing mechanism for kinase activity measurement.
摘要：
T4多核苷酸激酶(T4PNK)有助于DNA重组和修复。在这项工作中,创建了基于磷酸盐柱[5]芳烃@钯纳米颗粒@还原氧化石墨烯纳米复合材料（PP5@PdNPs@rGO）的电化学生物传感器，以识别T4PNK活性。用于复合甲苯胺蓝(TB)客体分子的PP5是水溶性的。使用T4PNK和ATP，底物DNA，其中包括一个5'-羟基，最初通过硫醇单元的化学吸附在金电极表面上自组装。强磷酸盐-Zr4+-磷酸盐化学允许Zr4+充当磷酸化DNA和PP5@PdNP@rGO之间的桥梁。通过超分子主客体识别连接，TB分子能够穿透PP5腔，它们产生了更强的电化学响应。检出限为5×10-7UmL-1，电化学信号在10-6至1UmL-1T4PNK浓度范围内呈线性关系。它在测量HeLa细胞裂解物相关的PNK活性和筛选PNK抑制剂方面也是有效的。核苷酸激酶靶向药物开发,临床诊断,和筛选抑制剂都将从建议的技术中受益匪浅，这为激酶活性测量提供了独特的传感机制。
