Investigation of C-terminal domain of SARS nucleocapsid protein–Duplex DNA interaction using transistors and binding-site models

Hsu, You-Ren; Kang, Yen-Wen; Fang, Jung-Ying; Lee, Geng-Yen; Chyi, Jen-Inn; Chang, Chung-ke; Huang, Chih-Cheng; Hsu, Chen-Pin; Huang, Tai-huang; Huang, Yu-Fen; Sun, Yuh-Chang; Hsu, Chia-Hsien; Chen, Chih-Chen; Li, Sheng-Shian; Yeh, J. Andrew; Yao, Da-Jeng; Ren, Fan; Wang, Yu-Lin

AlGaN/GaN high electron mobility transistors (HEMTs) were used to sense the binding between double stranded DNA (dsDNA) and the severe acute respiratory syndrome coronavirus (SARS-CoV) nucleocapsid protein (N protein). The sensing signals were the drain current change of the HEMTs induced by the protein–dsDNA binding. Binding-site models using surface coverage ratios were utilized to analyze the signals from the HEMT-based sensors to extract the dissociation constants and predict the number of binding sites. Two dissociation constants, K(D1) = 0.0955 nM, K(D2) = 51.23 nM, were obtained by fitting the experimental results into the two-binding-site model. The result shows that this technique is more competitive than isotope-labeling electrophoretic mobility shift assay (EMSA). We demonstrated that AlGaN/GaN HEMTs were highly potential in constructing a semiconductor-based-sensor binding assay to extract the dissociation constants of nucleotide–protein interaction.