Department of Chemistry , National Taiwan University , No. 1, Sec. 4, Roosevelt Road , Taipei 106 , Taiwan.
Genomics Research Center , Academia Sinica , No. 128, Academia Road, Sec. 2 , Nankang, Taipei 115 , Taiwan.
ACS Appl Mater Interfaces. 2018 Apr 18;10(15):12311-12316. doi: 10.1021/acsami.8b01917. Epub 2018 Apr 5.
A graphene field-effect transistor (G-FET) with the spacious planar graphene surface can provide a large-area interface with cell membranes to serve as a platform for the study of cell membrane-related protein interactions. In this study, a G-FET device paved with a supported lipid bilayer (referred to as SLB/G-FET) was first used to monitor the catalytic hydrolysis of the SLB by phospholipase D. With excellent detection sensitivity, this G-FET was also modified with a ganglioside G-enriched SLB (G-SLB/G-FET) to detect cholera toxin B. Finally, the GM1-SLB/G-FET was employed to monitor amyloid-beta 40 (Aβ40) aggregation. In the early nucleation stage of Aβ40 aggregation, while no fluorescence was detectable with traditional thioflavin T (ThT) assay, the prominent electrical signals probed by G-SLB/G-FET demonstrate that the G-FET detection is more sensitive than the ThT assay. The comprehensive kinetic information during the Aβ40 aggregation could be collected with a G-SLB/G-FET, especially covering the kinetics involved in the early stage of Aβ40 aggregation. These experimental results suggest that SLB/G-FETs hold great potential as a powerful biomimetic sensor for versatile investigations of membrane-related protein functions and interaction kinetics.
石墨烯场效应晶体管(G-FET)具有宽敞的平面石墨烯表面,可以提供与细胞膜大面积的接口,作为研究细胞膜相关蛋白相互作用的平台。在这项研究中,首先使用铺有支撑脂质双层(简称 SLB/G-FET)的 G-FET 器件来监测磷脂酶 D 对 SLB 的催化水解。这种 G-FET 具有出色的检测灵敏度,还可以通过神经节苷脂 G 富集的 SLB(G-SLB/G-FET)进行修饰,以检测霍乱毒素 B。最后,GM1-SLB/G-FET 被用于监测淀粉样蛋白-β40(Aβ40)聚集。在 Aβ40 聚集的早期成核阶段,传统的硫黄素 T(ThT)检测法无法检测到荧光,而 G-SLB/G-FET 探测到的显著电信号表明 G-FET 检测比 ThT 检测更灵敏。G-SLB/G-FET 可以收集 Aβ40 聚集过程中的综合动力学信息,特别是包括 Aβ40 聚集早期阶段的动力学信息。这些实验结果表明,SLB/G-FET 作为一种强大的仿生传感器,具有很大的潜力,可用于研究与膜相关的蛋白质功能和相互作用动力学。