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Using the QCM Biosensor-Based T7 Phage Display Combined with Bioinformatics Analysis for Target Identification of Bioactive Small Molecule

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Plant Chemical Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1795))

Abstract

Identification of target proteins that directly bind to bioactive small molecule is of great interest in terms of clarifying the mode of action of the small molecule as well as elucidating the biological phenomena at the molecular level. Of the experimental technologies available, T7 phage display allows comprehensive screening of small molecule-recognizing amino acid sequence from the peptide libraries displayed on the T7 phage capsid. Here, we describe the T7 phage display strategy that is combined with quartz-crystal microbalance (QCM) biosensor for affinity selection platform and bioinformatics analysis for small molecule-recognizing short peptides. This method dramatically enhances efficacy and throughput of the screening for small molecule-recognizing amino acid sequences without repeated rounds of selection. Subsequent execution of bioinformatics programs allows combinatorial and comprehensive target protein discovery of small molecules with its binding site, regardless of protein sample insolubility, instability, or inaccessibility of the fixed small molecules to internally located binding site on larger target proteins when conventional proteomics approaches are used.

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Acknowledgements

This work is partly supported by JSPS KAKENHI Grant Number 19380066 (F.S.), 19750145 (Y.T.), and 17K01363 (Y.T.).

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Authors and Affiliations

  1. Faculty of Science and Technology, Department of Applied Biological Science, Tokyo University of Science, Chiba, Japan

    Yoichi Takakusagi, Kaori Takakusagi, Fumio Sugawara & Kengo Sakaguchi

  2. Department of Molecular Imaging and Theranostics, National Institute of Radiological Sciences (NIRS), National Institutes for Quantum and Radiological Science and Technology (QST), Chiba, Japan

    Yoichi Takakusagi & Kaori Takakusagi

Authors
  1. Yoichi Takakusagi
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  2. Kaori Takakusagi
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  3. Fumio Sugawara
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  4. Kengo Sakaguchi
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Corresponding author

Correspondence to Yoichi Takakusagi .

Editor information

Editors and Affiliations

  1. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Friedrich Fauser

  2. Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA

    Martin Jonikas

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Takakusagi, Y., Takakusagi, K., Sugawara, F., Sakaguchi, K. (2018). Using the QCM Biosensor-Based T7 Phage Display Combined with Bioinformatics Analysis for Target Identification of Bioactive Small Molecule. In: Fauser, F., Jonikas, M. (eds) Plant Chemical Genomics. Methods in Molecular Biology, vol 1795. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7874-8_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7874-8_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7873-1

  • Online ISBN: 978-1-4939-7874-8

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