Receptor signal transduction protocols / edited by Gary B. Willars and R.A. John Challiss. — 3rd ed. — New York : Humana Press, 2011. – (58.17/M592/v.746)

Contents

    Contents
    
    preface
    contributors
    Part I DEVELOPING MODELS FOR STUDYING GPCRs
    1 Inducible Expression of G Protein-Coupled Receptors in Transfected cells
    2 Using the Flp-In T-Rex System to Regulate GPCR Expression
    3 Viral Infection for GPCR Expression in Eukaryotic Cells
    4 Generation of Epitope-Tagged GPCRs
    5 The Use of Site Directed Mutagenesis to Study GPCRs
    6 Approaches to Study GPCR Regulation in Native Systems
    7 Heterologous Expression of GPCRs in Fission Yeast
    PART II EXAMINING GPCR EXPRESSION AND AGONIST-INDUCED COVALENT MODIFICATIONS
    8 Radioligand Binding Methods for Membrane Preparations aim Intact Cells
    9 Quantification of GPCR mRNA Using Real-Time RT PCR
    10 Determining Allosteric Modulator Mechanism of Action: Integration of Radioligand Binding and Functional Assay Data
    11 Design and Use of Fluorescent Ligands to Study Ligand-Receptor Interactions in Single Living Cells
    12 Examining Site-Specific GPCR Phosphorylation
    13 Ubiquitination of GPCRs
    PART III EXAMINING EARLY EVENTS IN GPCR SIGNALING
    14 [35S]GTPγS Binding as an Index of Total G-Protein and Gα-Subtype-Specific Activation by GPCRs
    15 Using Calcium Imaging as a Readout of GPCR Activation
    16 Measuring Spatiotemporal Dynamics of Cyclic AMP Signaling in Real-Time Using FRET-Based Biosensors
    17 Determining the Activation of Rho as an Index of Receptor Coupling to G12/13 Proteins 317
    18 The Use of Translocating Fluorescent Biosensors for Real Time Monitoring of GPCR-Mediated Signaling Events
    PART IV RECEPTOR--RECEPTOR AND RECEPTOR--PROTEIN INTERACTIONS
    19 Study of GPCR-Protein Interactions Using Gel Overlay Assays and Glutathione-S-Transferase-Fusion Protein Pull-Downs
    20 Study of GPCR-Protein Interactions by BRET
    21 Time Resolved FRET Strategy with Fluorescent Ligands to Analyze Receptor Interactions in Native Tissues: Application to GPCR Oligomerization
    22 Peptide Affinity Purification for the Isolation and Identification of GPCR-Associated Protein Complexes
    23 Tandem Affinity Purification and Identification of GPCR-Associated Protein Complexes
    24 Identification of GPCR Localization in Detergent Resistant Membranes 411
    25 Analysis of GPCR Localization and Trafficking
    PART V STATISTICAL METHODS
    26 Statistical Methods in Research
    Index