Book Volume 1555 2017

SH2 Domains

Methods and Protocols

Editors:

ISBN: 978-1-4939-6760-5 (Print) 978-1-4939-6762-9 (Online)

Table of contents (31 protocols)

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  1. Front Matter

    Pages i-xiii

  2. Overview

    1. Front Matter

      Pages 1-1

    2. No Access

      Protocol

      Pages 3-35

      Introduction: History of SH2 Domains and Their Applications

    3. No Access

      Protocol

      Pages 37-43

      What Have We Learned from SH2 Domains?

  3. Computational Approaches

    1. Front Matter

      Pages 45-45

    2. No Access

      Protocol

      Pages 47-58

      Hidden Markov Models for Protein Domain Homology Identification and Analysis

    3. No Access

      Protocol

      Pages 59-75

      Classification and Lineage Tracing of SH2 Domains Throughout Eukaryotes

    4. No Access

      Protocol

      Pages 77-81

      SH2 Ligand Prediction–Guidance for In-Silico Screening

    5. No Access

      Protocol

      Pages 83-97

      An Efficient Semi-supervised Learning Approach to Predict SH2 Domain Mediated Interactions

    6. No Access

      Protocol

      Pages 99-113

      Proteomic Clustering Analysis of SH2 Domain Datasets

  4. Experimental Tools

    1. Front Matter

      Pages 115-115

    2. No Access

      Protocol

      Pages 117-162

      Expression and Production of SH2 Domain Proteins

    3. No Access

      Protocol

      Pages 163-172

      Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay

    4. No Access

      Protocol

      Pages 173-182

      Purification of SOCS (Suppressor of Cytokine Signaling) SH2 Domains for Structural and Functional Studies

    5. No Access

      Protocol

      Pages 183-198

      Expression and Purification of SH2 Domains Using Baculovirus Expression System

    6. No Access

      Protocol

      Pages 199-223

      Functionally Altered SH2 Domains for Biochemical Studies: Loss-of-Function Mutant and Domain Concatenation

    7. No Access

      Protocol

      Pages 225-254

      Creation of Phosphotyrosine Superbinders by Directed Evolution of an SH2 Domain

  5. Structural Analysis and Binding Assays

    1. Front Matter

      Pages 255-255

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      Protocol

      Pages 257-267

      Structural Characterization of Monomeric/Dimeric State of p59fyn SH2 Domain

    3. No Access

      Protocol

      Pages 269-290

      NMR Chemical Shift Mapping of SH2 Peptide Interactions

    4. No Access

      Protocol

      Pages 291-305

      Calorimetric Measurement of SH2 Domain Ligand Affinities

    5. No Access

      Protocol

      Pages 307-330

      Binding Assays Using Recombinant SH2 Domains: Far-Western, Pull-Down, and Fluorescence Polarization

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      Protocol

      Pages 331-347

      In-Solution SH2 Domain Binding Assay Based on Proximity Ligation

  6. High-Throughput Screening and Proteomics

    1. Front Matter

      Pages 349-349

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      Protocol

      Pages 351-356

      Alpha-Based Multiplexed Assay for Identifying SH2 Domain Antagonists

    3. No Access

      Protocol

      Pages 357-373

      Characterizing SH2 Domain Specificity and Network Interactions Using SPOT Peptide Arrays

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