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Engineering SH2 Domains with Tailored Specificities and Affinities

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SH2 Domains

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2705))

Abstract

The Src Homology 2 (SH2) domain is an emerging biotechnology with applications in basic science, drug discovery, and even diagnostics. The SH2 domains rapid uptake into different areas of research is a direct result of the wealth of information generated on its biochemical, biological, and biophysical role in mammalian cell biology. Functionally, the SH2 domain binds and recognizes specific phosphotyrosine (pTyr) residues in the cell to mediate protein–protein interactions (PPIs) that govern signal transduction networks. These signal transduction networks are responsible for relaying growth and stress state signals to the cell’s nucleus, ultimately effecting a change in cell biology. Protein engineers have been able to increase the affinity of SH2 domains for pTyr while also tailoring the domains’ specificity to unique amino acid sequences flanking the pTyr residue. In this way, it has been possible to develop unique SH2 variants for use in affinity-purification coupled to mass spectrometry (AP-MS) experiments, microscopy, or even synthetic biology. This chapter outlines methods to tailor the affinity and specificity of virtually any human SH2 domain using a combination of rational engineering and phage-display approaches.

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Abbreviations

PTM:

Posttranslational modification

pTyr:

Phosphotyrosine

SH2:

Src-Homology 2

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Author information

Authors and Affiliations

  1. Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada

    Gregory D. Martyn, Gianluca Veggiani & Sachdev S. Sidhu

  2. Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada

    Gregory D. Martyn & Sachdev S. Sidhu

Authors
  1. Gregory D. Martyn
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  2. Gianluca Veggiani
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  3. Sachdev S. Sidhu
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Corresponding author

Correspondence to Sachdev S. Sidhu .

Editor information

Editors and Affiliations

  1. School of Biosciences, College of Life and Environmental Sciences, University of Birmingham, Birmingham, UK

    Teresa Carlomagno

  2. Signalling Research Centres BIOSS &CIBSS, University of Freiburg, Freiburg, Germany

    Maja Köhn

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Martyn, G.D., Veggiani, G., Sidhu, S.S. (2023). Engineering SH2 Domains with Tailored Specificities and Affinities. In: Carlomagno, T., Köhn, M. (eds) SH2 Domains. Methods in Molecular Biology, vol 2705. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3393-9_17

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  • DOI: https://doi.org/10.1007/978-1-0716-3393-9_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3392-2

  • Online ISBN: 978-1-0716-3393-9

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