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A Novel Method for Determining the Kinetics of G Protein-Coupled Receptor Plasma Membrane Expression

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A new approach for characterizing the plasma membrane delivery of G protein-coupled receptors is described in this chapter. This approach uses an existing technology, the regulated secretion/aggregation technology (RPD ), to cause the accumulation of G protein-coupled receptors in the ER of cells. The trafficking of accumulated receptor from the ER to the plasma membrane of cells can be initiated by a small, membrane permeable ligand. The plasma membrane delivery of receptors can be monitored using radioligand binding or microscopy or both techniques. Using this new approach it should be possible to determine the rate of plasma membrane delivery of a broad range of G protein-coupled receptors and thus facilitate learning more about their anterograde trafficking mechanisms.
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