Identification of preferred protein interactions by phage-display of the human Src homology-3 proteome
| Autor: | Kalle Saksela, Marika Vähä-Jaakkola, G. Herman Renkema, Iivari Kleino, Jing-Huan Wang, Michael Liss, Satu Kärkkäinen, Marita Hiipakka, Ralf Wagner |
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| Rok vydání: | 2005 |
| Předmět: |
Phage display
Proteome Recombinant Fusion Proteins Ubiquitin-Protein Ligases Genetic Vectors Scientific Report Vesicular Transport Proteins Computational biology Plasma protein binding Biology Protein Serine-Threonine Kinases Ligands Biochemistry SH3 domain Gene Products nef Protein–protein interaction src Homology Domains 03 medical and health sciences Peptide Library Genetics Humans Peptide library Molecular Biology 030304 developmental biology Glutathione Transferase 0303 health sciences 030302 biochemistry & molecular biology Signal transducing adaptor protein Membrane Proteins Proteins 3. Good health Sorting nexin ADAM Proteins p21-Activated Kinases Carrier Proteins Protein Binding |
| Zdroj: | EMBO reports. 7(2) |
| ISSN: | 1469-221X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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