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Alpha-Based Multiplexed Assay for Identifying SH2 Domain Antagonists

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SH2 Domains

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1555))

Abstract

Constitutive activation of STAT3/5b frequently occurs in various human malignancies. STAT3/5b activation involves dimerization via intermolecular pTyr-SH2 binding; therefore, antagonizing this interaction is a feasible approach to inhibit STAT3/5b activation for cancer therapy. We have developed a multiplexed assay to assess STAT3- and STAT5b-SH2 binding in a single well by combining AlphaLISA and AlphaScreen beads. In this chapter, we describe application of the method for the purpose of identifying new STAT3 and STAT5b antagonists.

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Acknowledgment

This work was supported by National Institute of Biomedical Innovation (NIBO) grant number 06–02, JSPS KAKENHI Grant Number 23510281/26430166, and Pharma Valley Center (PVC) Shizuoka, Japan.

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Authors and Affiliations

  1. Center for Drug Discovery, Graduate School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka, 422-8526, Japan

    Akira Asai & Kazuyuki Takakuma

  2. Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Saitama, Japan

    Kazuyuki Takakuma

Authors
  1. Akira Asai
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  2. Kazuyuki Takakuma
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Corresponding author

Correspondence to Akira Asai .

Editor information

Editors and Affiliations

  1. Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, Farmington, Connecticut, USA

    Kazuya Machida

  2. Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA

    Bernard A. Liu

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Asai, A., Takakuma, K. (2017). Alpha-Based Multiplexed Assay for Identifying SH2 Domain Antagonists. In: Machida, K., Liu, B. (eds) SH2 Domains. Methods in Molecular Biology, vol 1555. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6762-9_19

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  • DOI: https://doi.org/10.1007/978-1-4939-6762-9_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6760-5

  • Online ISBN: 978-1-4939-6762-9

  • eBook Packages: Springer Protocols

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