Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Korea.
Department of Rural System Engineering, Seoul National University, Seoul 08826, Korea.
Sensors (Basel). 2018 Jan 5;18(1):136. doi: 10.3390/s18010136.
An immobilized liposome electrode (ILE)-based sensor was developed to quantify conformational changes of the proteins under various stress conditions. The ILE surface was characterized by using a tapping-mode atomic force microscopy (TM-AFM) to confirm surface immobilization of liposome. The uniform layer of liposome was formed on the electrode. The current deviations generated based on the status of the proteins under different stress were then measured. Bovine carbonic anhydrase (CAB) and lysozyme were tested with three different conditions: native, reduced and partially denatured. For both proteins, a linear dynamic range formed between denatured concentrations and output electric current signals was able to quantify conformational changes of the proteins. The pattern recognition (PARC) technique was integrated with ILE-based sensor to perform data analysis and provided an effective method to improve the prediction of protein structural changes. The ILE-based stress sensor showed potential of leveraging the amperometric technique to manifest activity of proteins based on various external conditions.
一种固定化脂质体电极（ILE）传感器被开发出来，以定量研究蛋白质在各种应激条件下的构象变化。通过使用原子力显微镜的轻敲模式（TM-AFM）对 ILE 表面进行了特征描述，以确认脂质体的表面固定化。在电极上形成了均匀的脂质体层。然后测量基于不同应激状态下蛋白质状态产生的电流偏差。用三种不同的条件测试了牛碳酸酐酶（CAB）和溶菌酶：天然、还原和部分变性。对于这两种蛋白质，在变性浓度和输出电流信号之间形成了线性动态范围，能够定量蛋白质的构象变化。模式识别（PARC）技术与基于 ILE 的传感器集成在一起进行数据分析，为预测蛋白质结构变化提供了一种有效的方法。ILE 基应激传感器显示出利用安培技术根据各种外部条件表现蛋白质活性的潜力。
