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

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

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2 Citations (Scopus)

Abstract

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 = 0.0955 nM, KD2 = 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.

Original languageEnglish
Pages (from-to)334-339
Number of pages6
JournalSensors and Actuators, B: Chemical
Volume193
DOIs
Publication statusPublished - 2014 Mar 31

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Keywords

  • Binding sites
  • Dissociation constants
  • GaN
  • HEMTs
  • SARS
  • Sensors

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Instrumentation
  • Condensed Matter Physics
  • Surfaces, Coatings and Films
  • Metals and Alloys
  • Electrical and Electronic Engineering
  • Materials Chemistry

Cite this

Hsu, Y. R., Kang, Y. W., Fang, J. Y., Lee, G. Y., Chyi, J. I., Chang, C. K., Huang, C. C., Hsu, C. P., Huang, T. H., Huang, Y. F., Sun, Y. C., Hsu, C. H., Chen, C. C., Li, S. S., Yeh, J. A., Yao, D. J., Ren, F., & Wang, Y. L. (2014). Investigation of C-terminal domain of SARS nucleocapsid protein-Duplex DNA interaction using transistors and binding-site models. Sensors and Actuators, B: Chemical, 193, 334-339. https://doi.org/10.1016/j.snb.2013.11.087