College of Chemistry and Material Science, Food Safety Analysis and Test Engineering Technology Research Center of Shandong Province, Shandong Agricultural University, Tai'an, Shandong 271018, PR China.
Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, College of Science, Nanjing Forestry University, Nanjing 210037, PR China.
Anal Chem. 2024 Aug 13;96(32):13086-13095. doi: 10.1021/acs.analchem.4c01610. Epub 2024 Jul 29.
The OPECT biosensing platform, which connects optoelectronics and biological systems, offers significant amplification and more possibilities for research in biological applications. In this work, a homogeneous organic photoelectrochemical transistor (OPECT) biosensor based on a BiS/BiMoO heterojunction was constructed to detect METTL3/METTL14 protein activity. The METTL3/METTL14 complex enzyme was used to catalyze adenine (A) on an RNA strand to mA, protecting mA-RNA from being cleaved by an toxin (MazF). Alkaline phosphatase (ALP) catalyzed the conversion of NaSPO to HS through an enzymatic reaction. Due to the adoption of the strategy of no fixation on the electrode, the generated HS was easy to diffuse to the surface of the ITO electrode. The BiS/BiMoO heterojunction was formed in situ through a chemical replacement reaction with BiMoO, improving photoelectric conversion efficiency and realizing signal amplification. Based on this "signal on" mode, METTL3/METTL14 exhibited a wide linear range (0.00001-25 ng/μL) between protein concentration and photocurrent intensity with a limit of detection (LOD) of 7.8 fg/μL under optimal experimental conditions. The applicability of the developed method was evaluated by investigating the effect of four plasticizers on the activity of the METTL3/METTL14 protein, and the molecular modeling technique was employed to investigate the interaction between plasticizers and the protein.
OPECT 生物传感平台将光电和生物系统连接起来，为生物应用研究提供了显著的放大和更多的可能性。在这项工作中，构建了一种基于 BiS/BiMoO 异质结的均相有机光电化学晶体管（OPECT）生物传感器，用于检测 METTL3/METTL14 蛋白活性。METTL3/METTL14 复合酶用于催化 RNA 链上的腺嘌呤（A）转变为 mA，从而保护 mA-RNA 免受毒素（MazF）的切割。碱性磷酸酶（ALP）通过酶促反应将 NaSPO 转化为 HS。由于采用了不在电极上固定的策略，生成的 HS 很容易扩散到 ITO 电极表面。BiS/BiMoO 异质结通过与 BiMoO 的化学取代反应原位形成，提高了光电转换效率并实现了信号放大。基于这种“信号开启”模式，在最佳实验条件下，METTL3/METTL14 的蛋白浓度和光电流强度之间表现出宽的线性范围（0.00001-25 ng/μL），检测限（LOD）为 7.8 fg/μL。通过研究四种增塑剂对 METTL3/METTL14 蛋白活性的影响，评估了所开发方法的适用性，并采用分子建模技术研究了增塑剂与蛋白质之间的相互作用。
