First Author | Sparks AB | Year | 1996 |
Journal | Nat Biotechnol | Volume | 14 |
Issue | 6 | Pages | 741-4 |
PubMed ID | 9630982 | Mgi Jnum | J:35786 |
Mgi Id | MGI:83231 | Doi | 10.1038/nbt0696-741 |
Citation | Sparks AB, et al. (1996) Cloning of ligand targets: systematic isolation of SH3 domain-containing proteins. Nat Biotechnol 14(6):741-4 |
abstractText | 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. |