Signal Transduction Protocols

Refining Efficacy: Allosterism and Bias in G Protein-Coupled Receptor Signaling.- Imaging-based Approaches to Understanding G Protein-Coupled Receptor Signaling Complexes.- Improving Drug Discovery with Contextual Assays and Cellular Systems Analysis.- RGS-insensitive Ga Subunits: Probes of Ga Subtype-selective Signaling and Physiological Functions of RGS Proteins.- Bioinformatic Approaches to Metabolic Pathways Analysis.- Studying Ligand Efficacy at G Protein-Coupled Receptors Using FRET.- Using BRET to Detect Ligand-Specific Conformational Changes in Preformed Signaling Complexes.- Reconstitution of G Protein-Coupled Receptors into a Model Bilayer System: Reconstituted High Density Lipoprotein Particles (rHDL).- Using Quantitative BRET to Assess G Protein-Coupled Receptor Homo- and Hetero-Dimerization.- Cell-Surface Protein-Protein Interaction Analysis with Time-Resolved FRET and Snap-tag Technologies: Application to G Protein-Coupled Receptor Oligomerization.- Analysis of GPCR/Ion Channel Interactions.- Multicolor BiFC Analysis of G Protein bg Complex Formation and Localization.- Real-Time BRET Assays to Measure G Protein/Effector Interactions.- Luminescent Biosensors for Real-Time Monitoring of Intracellular cAMP.- Simultaneous Real Time Imaging of Signal Oscillations Using Multiple Fluorescence-Based Reporters.- Using FRET-based Reporters to Visualize Subcellular Dynamics of Protein Kinase A Activity.- Genetically Encoded Fluorescent Reporters to Visualize Protein Kinase C Activation in Live Cells.- Visualizing Receptor Endocytosis and Trafficking.- Investigating G Protein-Coupled Receptor Endocytosis and Trafficking by TIR-FM.- Visualizing G Protein-Coupled Receptor Signalsomes Using Confocal Immunofluorescence Microscopy.- Detection and Characterization of Receptor Interactions with PDZ Domains.- Tandem Affinity Purification and Identification of Heterotrimeric G-Protein Associated Proteins.- Study of G Protein-Coupled Receptor/ßarrestinInteractions within Endosomes Using FRAP.- Disrupting Protein Complexes Using Tat-tagged Peptide Mimics.- Protein-fragment Complementation Assays for Large-Scale Analysis, Functional Dissection and Dynamic Studies of Protein-Protein Interactions in Living Cells.