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

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 pro...

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Bibliographic Details
Published in:Sensors and actuators. B, Chemical Chemical, 2014-03, Vol.193, p.334-339
Main Authors: 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
Format: Article
Language:English
Subjects:
Aluminum gallium nitrides
Assaying
Binding
Binding sites
Coronaviridae
Deoxyribonucleic acid
Dissociation constants
Gallium nitrides
GaN
HEMTs
High electron mobility transistors
SARS
SARS coronavirus
Semiconductor devices
Sensors
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Summary: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, KD1=0.0955nM, KD2=51.23nM, 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.
ISSN:0925-4005
1873-3077
0925-4005
DOI:10.1016/j.snb.2013.11.087