Receptor-receptor interactions [electronic resource] / edited by P. Michael Conn.

Front Cover; Receptor-Receptor Interactions; Copyright; Contents; Contributors; Preface; References; Chapter 1: Spatial Intensity Distribution Analysis (SpIDA) : A New Tool for Receptor Tyrosine Kinase Activation and Transactivation Quantification; Introduction; 1.1. Theory of Spatial Intensity Distribution Analysis; 1.1.1. Theoretical basis of SpIDA; 1.2. SpIDA: Examples of Application to RTK; 1.2.1. Quantification of EGFR-eGFP; 1.2.2. Application to RTK transactivation in a native system: neurons expressing dopamine receptors; 1.3. Procedure for SpIDA; 1.3.1. Material and apparatus.

1.3.1.1. Sample preparation; 1.3.2. Necessary experimental controls : defining the QB for monomeric moiety; 1.3.2.1. Determining the monomeric QB for receptors tagged with fluorescent proteins; 1.3.2.2. Determining the monomeric QB for antibody labeling using detection of a reference monomeric protein; 1.3.2.3. Determining the monomeric QB by using pharmacological agents that block oligomerization and induce monomeric conformation; 1.3.3. Image acquisitions; 1.3.4. Analysis with the SpIDA program graphical user interface; 1.3.4.1. Description of histogram parameters; 1.3.4.2. SpIDA GUI procedures.

1.3.5. Determination of analog detector signal broadening; 1.3.5.1. Analog detector calibration procedure; 1.3.6. Data interpretation and pharmacological analysis; 1.4. Discussion; Acknowledgments; References; Chapter 2: Dimerization of Nuclear Receptors; Introduction; 2.1. Methods; 2.1.1. Studies of NR-NR interactions through protein crystallization; 2.1.1.1. Required materials; 2.1.1.2. Protocol; 2.1.2. Expression and purification of NR-NR complexes; 2.1.2.1. Required materials; 2.1.2.2. Protocol; 2.1.3. Monitoring NR-NR interactions by noncovalent electrospray ionization mass spectrometry.

2.1.3.1. Required materials2.1.3.2. Protocol; 2.1.4. Monitoring NR-NR interactions by electrophoretic mobility shift assays; 2.1.4.1. Required materials; 2.1.4.2. Protocol; 2.1.4.2.1. In vitro transcription-translation; 2.1.4.2.2. Electrophoretic mobility shift assay; 2.1.5. Two-hybrid assays to define NR-NR interactions in living cells; 2.1.5.1. Required materials; 2.1.5.2. Protocol; 2.1.5.2.1. Transient transfection of HeLa cells; 2.1.5.2.2. Cell lysis; 2.1.5.2.3. Preparation of luciferase assay; 2.1.5.2.4. Luciferase measurement; 2.1.5.2.5. Measurement of {Beta]gal activity.

2.1.5.2.6. Normalization; 2.1.6. Fluorescence cross-correlation spectroscopy to measure the concentrations and interactions of NRs in living cells; 2.1.6.1. Required materials; 2.1.6.2. Protocol; 2.1.6.2.1. Cell culture and transfection; 2.1.6.2.2. Fluorescence cross-correlation microscopy; 2.1.6.2.3. Data analysis; Acknowledgments; References; Chapter 3 : Network Analysis to Uncover the Structural Communication in GPCRs; Introduction; 3.1. Materials; 3.2. Methods; 3.2.1. Workflow of the PSN-MD and PSN-ENM approaches; 3.2.2. Building the PSG; 3.2.3. Search for the shortest communication paths.

3.3. Discussion.

This new volume of Methods in Cell Biology looks at receptor-receptor interactions, with sections on allosteric and effector interactions, crystallization and modeling, measuring receptor-receptor interactions and oligomerization in individual classes. With cutting-edge material, this comprehensive collection is intended to guide researchers of receptor-receptor interactions for years to come. Covers sections on allosteric and effector interactions, crystallization and modeling, measuring receptor-receptor interactions and oligomerization in individual class.

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