title: NMR Chemical Shift Mapping of SH2 Peptide Interactions.
abstract: Heteronuclear single quantum coherence (HSQC) nuclear magnetic resonance (NMR) experiments offer a rapid and high resolution approach to gaining binding and conformational insights into a protein-peptide interaction. By tracking1H and15N chemical shift changes over the course of a peptide titration into isotopically labeled protein, amide NH pairs of amino acids whose chemical environment changes upon peptide binding can be identified. When mapped onto a structure of the protein, this approach can identify the peptide-binding interface or regions undergoing conformation changes within a protein upon ligand binding. Monitoring NMR chemical shift changes can also serve as a screening technique to identify novel interaction partners for a protein or to determine the binding affinity of a weak protein-peptide interaction. Here, we describe the application of NMR chemical shift mapping to the study of peptide binding to the C-terminal SH2 domain of PLCγ1.
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, 215 UCB, Boulder, CO, 80309, USA.
authors.forename: Marissa A
authors.initials: MA
authors.lastname: McKercher
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, 215 UCB, Boulder, CO, 80309, USA. deborah.wuttke@colorado.edu.
authors.forename: Deborah S
authors.initials: DS
authors.lastname: Wuttke
mesh_terms: D000465:Algorithms; D000595:Amino Acid Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D015870:Gene Expression; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D018909:src Homology Domains
chemical_list: D010455:Peptides; D011993:Recombinant Fusion Proteins
keywords: Chemical shift mapping (CSM); Nuclear magnetic resonance (NMR); PLCγ1; Peptide; Phosphotyrosine; SH2 domain