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SH2 Domains pp 163–172Cite as

Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay

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

Abstract

When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.

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References

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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). Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay. 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_9

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  • DOI: https://doi.org/10.1007/978-1-4939-6762-9_9

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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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