title: Cloning of ligand targets: systematic isolation of SH3 domain-containing proteins.
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.
authors.affiliation: Department of Biology, University of North Carolina, Chapel Hill 27599, USA.
authors.forename: A B
authors.initials: AB
authors.lastname: Sparks
authors.forename: N G
authors.initials: NG
authors.lastname: Hoffman
authors.forename: S J
authors.initials: SJ
authors.lastname: McConnell
authors.forename: D M
authors.initials: DM
authors.lastname: Fowlkes
authors.forename: B K
authors.initials: BK
authors.lastname: Kay
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D006801:Humans; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018909:src Homology Domains
chemical_list: D008024:Ligands; D011506:Proteins