title: A repertoire library that allows the selection of synthetic SH2s with altered binding specificities.
abstract: Tyrosine phosphorylation is one of the major mechanisms involved in the intracellular propagation of external signals. Strategies aimed at interfering with this process might allow the control of several cellular phenotypes. SH2 domains mediate protein-protein interactions by recognizing phosphotyrosine (pY) residues in the context of specific phosphopeptides. We created an SH2-scaffolded repertoire library by randomly mutagenizing five critical amino acid positions in the specificity-determining region of the PLCgamma C-terminal SH2 domain. Synthetic SH2 domains were selected from the library using biotinylated phosphopeptides derived from a natural PLCgamma-SH2 ligand as well as unrelated SH2 ligands. The isolated SH2s displayed high binding affinity constants for the selecting peptides and were capable of interacting with the corresponding proteins.
authors.affiliation: Department of Experimental Oncology, European Institute of Oncology, Via Ripamonti 435, 20141 Milan, Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Malabarba
authors.forename: E
authors.initials: E
authors.lastname: Milia
authors.forename: M
authors.initials: M
authors.lastname: Faretta
authors.forename: R
authors.initials: R
authors.lastname: Zamponi
authors.forename: P G
authors.initials: PG
authors.lastname: Pelicci
authors.forename: P P
authors.initials: PP
authors.lastname: Di Fiore
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D016296:Mutagenesis; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011485:Protein Binding; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D020349:Surface Plasmon Resonance; D013997:Time Factors; D020798:Two-Hybrid System Techniques; D014443:Tyrosine / Q000378:metabolism; D018909:src Homology Domains
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D019000:Phosphotyrosine; D014443:Tyrosine