SH2 Domains

Kazuya Machida, MD, PhD Dept of Genetics and Developmental Biology Univ of Connecticut Health Center Cell and Genome Sciences, Farmington, CT, USA Tel: 860-679-1830; Email: machida@neuron.uchc.edu

Introduction: History of SH2 Domains and Their Applications.- What Have We Learned from SH2 Domains?.- Hidden Markov Models for Protein Domain Homology Identification and Analysis.- Classification and Lineage Tracing of SH2 Domains Throughout Eukaryotes.- SH2 Ligand Prediction-Guidance for In-Silico Screening.- An Efficient Semi-Supervised Learning Approach to Predict SH2 Domain Mediated Interactions.- Proteomic Clustering Analysis of SH2 Domain Datasets.- Expression and Production of SH2 Domain Proteins.- Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.- Purification of SOCS (Suppressors of Cytokine Signalling) SH2 Domains for Structural and Functional Studies.- Expression and Purification of SH2 Domains Using Baculovirus Expression System.- Functionally Altered SH2 Domains for Biochemical Studies: Loss of Function Mutant and Domain Concatenation.- Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain.- Structural Characterization of Monomeric/Dimeric State of p59fyn SH2 Domain.- NMR Chemical Shift Mapping of SH2 Peptide Interactions.- Calorimetric Measurement of SH2 Domain Ligand Affinities.- Binding Assays Using Recombinant SH2 Domains: Far-Western, Pull Down, and Fluorescence Polarization.- In-Solution SH2 Domain Binding Assay Based on Proximity Ligation.- Alpha-Based Multiplexed Assay for Identifying SH2 Domain Antagonists.- Characterizing SH2 Domain Specificity and Network Interactions Using SPOT Peptide Arrays.- High Throughput Quantification of SH2 Domain-Phosphopeptide Interactions with Cellulose Peptide Conjugate Microarrays.- SH2 Domains as Affinity Reagents for Phosphotyrosine Protein Enrichment and Proteomic Analysis.- Identification of Tyrosine Phosphorylated Proteins by SH2 Domain Affinity Purification and Mass Spectrometry.- Analysis of the Global Changes in SH2 Binding Properties Using Mass Spectrometry Supported by Quantitative Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) Technique.- Using Reciprocal Protein-Peptide Array Screening to Unravel Protein Interaction Networks.- Rosette Assay: Highly Customizable Dot-Blot for SH2 Domain Screening.- Microwestern Arrays for Systems-Level Analysis of SH2 Domain-Containing Proteins.- SH2 Binding Site Protection Assay: A Method for Identification of SH2 Domain Interaction Partners by Exploiting SH2 Mediated Phosphosite Protection.- Real Time Single Molecule Visualization of SH2 Domain Membrane Recruitment in Growth Factor Stimulated Cells.- SH2 Domain-Based FRET Biosensor for Measuring BCR-ABL Activity in Living CML Cells.- SH2 Domain Histochemistry.