Improved silicon nanowire field-effect transistors for fast protein-protein interaction screening

Ti Yu Lin, Bor-Ran Li, Sheng Ta Tsai, Chien Wei Chen, Chung Hsuan Chen, Yit Tsong Chen*, Chien Yuan Pan

*Corresponding author for this work

Research output: Contribution to journalArticle

20 Scopus citations

Abstract

Understanding how proteins interact with each other is the basis for studying the biological mechanisms behind various physiological activities. Silicon nanowire field-effect transistors (SiNW-FETs) are sensitive sensors used to detect biomolecular interactions in real-time. However, the majority of the applications that use SiNW-FETs are for known interactions between different molecules. To explore the capability of SiNW-FETs as fast screening devices to identify unknown interacting molecules, we applied mass spectrometry (MS) to analyze molecules reversibly bound to the SiNW-FETs. Calmodulin (CaM) is a Ca2+-sensing protein that is ubiquitously expressed in cells and its interaction with target molecules is Ca2+-dependent. By modifying the SiNW-FET surface with glutathione, glutathione S-transferase (GST)-tagged CaM binds reversibly to the SiNW-FET. We first verified the Ca2+- dependent interaction between GST-CaM and purified troponin I, which is involved in muscle contraction, through the conductance changes of the SiNW-FET. Furthermore, the cell lysate containing overexpressed Ca2+/CaM- dependent protein kinase IIα induced a conductance change in the GST-CaM-modified SiNW-FET. The bound proteins were eluted and subsequently identified by MS as CaM and kinase. In another example, candidate proteins from neuronal cell lysates interacting with calneuron I (CalnI), a CaM-like protein, were captured with a GST-CalnI-modified SiNW-FET. The proteins that interacted with CalnI were eluted and verified by MS. The Ca2+-dependent interaction between GST-CalnI and one of the candidates, heat shock protein 70, was re-confirmed via the SiNW-FET measurement. Our results demonstrate the effectiveness of combining MS with SiNW-FETs to quickly screen interacting molecules from cell lysates.

Original languageEnglish
Pages (from-to)676-684
Number of pages9
JournalLab on a Chip
Volume13
Issue number4
DOIs
StatePublished - 21 Feb 2013

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    Lin, T. Y., Li, B-R., Tsai, S. T., Chen, C. W., Chen, C. H., Chen, Y. T., & Pan, C. Y. (2013). Improved silicon nanowire field-effect transistors for fast protein-protein interaction screening. Lab on a Chip, 13(4), 676-684. https://doi.org/10.1039/c2lc40772h