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Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome.

ABSTRACT: We have determined the human genome to contain 296 different Src homology-3 (SH3) domains and cloned them into a phage-display vector. This provided a powerful and unbiased system for simultaneous assaying of the complete human SH3 proteome for the strongest binding to target proteins of interest, without the limitations posed by short linear peptide ligands or confounding variables of more indirect methods for protein interaction screening. Studies involving three ligand proteins, human immunodeficiency virus-1 Nef, p21-activated kinase (PAK)2 and ADAM15, showed previously reported as well as novel SH3 partners with nanomolar affinities specific for them. This argues that SH3 domains may have a more dominant role in directing cellular protein interactions than has been assumed. Besides showing potentially important new SH3-directed interactions, these studies also led to the discovery of novel signalling proteins, such as the PAK2-binding adaptor protein POSH2 and the ADAM15-binding sorting nexin family member SNX30.

SUBMITTER: Karkkainen S

PROVIDER: S-EPMC1369250 | biostudies-literature | 2006 Feb

REPOSITORIES: biostudies-literature

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Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome.

Kärkkäinen Satu S Hiipakka Marita M Wang Jing-Huan JH Kleino Iivari I Vähä-Jaakkola Marika M Renkema G Herma GH Liss Michael M Wagner Ralf R Saksela Kalle K

EMBO reports 20060201 2

We have determined the human genome to contain 296 different Src homology-3 (SH3) domains and cloned them into a phage-display vector. This provided a powerful and unbiased system for simultaneous assaying of the complete human SH3 proteome for the strongest binding to target proteins of interest, without the limitations posed by short linear peptide ligands or confounding variables of more indirect methods for protein interaction screening. Studies involving three ligand proteins, human immunod ...[more]

PMID: 16374509

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Dataset Information

Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome.

Publications

Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome.

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