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Cloning of ligand targets: Systematic isolation of SH3 domain-containing proteins

Nature Biotechnology volume 14, pages 741–744 (1996)Cite this article

Abstract

Based on the prevalence of modular protein domains, such as Src homology domain 3 and 2 (SH3 and SH2), among important signaling molecules, we have sought to identify new SH3 domain-containing proteins. However, modest sequence similarity among these domains restricts the use of DNA-based methods for this purpose. To circumvent this limitation, we have developed a functional screen that permits the rapid cloning of modular domains based on their ligand-binding activity. Using operationally defined SH3 ligands from combinatorial peptide libraries, we screened a series of mouse and human cDNA expression libraries. We found that 69 of the 74 clones isolated encode at least one SH3 domain. These clones encode 18 different SH3-containing proteins, 10 of which have not been described previously. The isolation of entire repertoires of modular domain-containing proteins will prove invaluable in genome analysis and in bringing new targets into drug discovery programs.

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Author information

Author notes

  1. Brian K. Kay: briankay@email.unc.edu

Authors and Affiliations

  1. Curriculum in Genetics & Molecular Biology, University of North Carolina, Chapel Hill, NC, 27599

    Andrew B. Sparks

  2. Department of Biology, University of North Carolina, Chapel Hill, NC, 27599

    Noah G. Hoffman & Brian K. Kay

  3. Cytogen Corporation, Princeton, NJ, 08540

    Stephen J. McConnell & Dana M. Fowlkes

  4. Department of Pathology, University of North Carolina, Chapel Hill, NC, 27599

    Dana M. Fowlkes

  5. Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC, 27599

    Brian K. Kay

Authors
  1. Andrew B. Sparks
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  2. Noah G. Hoffman
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  3. Stephen J. McConnell
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  4. Dana M. Fowlkes
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  5. Brian K. Kay
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Sparks, A., Hoffman, N., McConnell, S. et al. Cloning of ligand targets: Systematic isolation of SH3 domain-containing proteins. Nat Biotechnol 14, 741–744 (1996). https://doi.org/10.1038/nbt0696-741

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  • DOI: https://doi.org/10.1038/nbt0696-741

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