title: Characterizing SH2 Domain Specificity and Network Interactions Using SPOT Peptide Arrays.
abstract: Src Homology 2 (SH2) domains are protein interaction modules that recognize and bind tyrosine phosphorylated ligands. Their ability to distinguish binding to over thousands of potential phosphotyrosine (pTyr) ligands within the cell is critical for the fidelity of receptor tyrosine kinase (RTK) signaling. Within humans there are over a hundred SH2 domains with more than several thousand potential ligands across many cell types and cell states. Therefore, defining the specificity of individual SH2 domains is critical for predicting and identifying their physiological ligands. Here, in this chapter, I describe the broad use of SPOT peptide arrays for examining SH2 domain specificity. An orientated peptide array library (OPAL) approach can uncover both favorable and non-favorable residues, thus providing an in-depth analysis to SH2 specificity. Moreover, I discuss the application of SPOT arrays for paneling SH2 ligand binding with physiological peptides.
authors.affiliation: Broad Institute of MIT and Harvard, 415 Main St., 5175 JJ, Cambridge, MA, 02142, USA. baliu@broadinstitute.org.
authors.forename: Bernard A
authors.initials: BA
authors.lastname: Liu
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D057166:High-Throughput Screening Assays; D008024:Ligands; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D010748:Phosphopeptides / Q000737:chemistry / Q000378:metabolism; D019000:Phosphotyrosine / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D012680:Sensitivity and Specificity; D018909:src Homology Domains
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides; D010748:Phosphopeptides; D019000:Phosphotyrosine
keywords: Oriented Peptide Array Library (OPAL); Phosphotyrosine (pTyr); SPOT peptide arrays; Specificity; Src Homology 2 (SH2)