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  • Title: Cloning of ligand targets: systematic isolation of SH3 domain-containing proteins.
    Author: Sparks AB, Hoffman NG, McConnell SJ, Fowlkes DM, Kay BK.
    Journal: Nat Biotechnol; 1996 Jun; 14(6):741-4. PubMed ID: 9630982.
    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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