Systematic and quantitative analysis of G protein-coupled receptor trafficking motifs.
Plasma membrane expression of G protein-coupled receptors (GPCRs) is a dynamic process balancing anterograde and retrograde trafficking. Multiple interrelated cellular processes determine the final level of cell surface expression, including endoplasmic reticulum (ER) export/retention, receptor internalization, recycling, and degradation. These processes are highly regulated to achieve specific localization to subcellular domains (e.g., dendrites or basolateral membranes) and to affect receptor signaling. Analysis of potential ER trafficking motifs within GPCRs requires careful consideration of intracellular dynamics, such as protein folding, ER export and retention, and glycosylation. This chapter presents an approach and methods for qualitative and quantitative assessment of these processes to aid in accurate identification of GPCR trafficking motifs, utilizing the analysis of a hydrophobic extracellular trafficking motif in α2C adrenergic receptors as a model system.
Department of Anesthesia/CCM, Stanford University Medical School, Stanford, California, USA.
Carl M
CM
Hurt
Vincent K
VK
Ho
Timothy
T
Angelotti
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique / Q000379:methods; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D021381:Protein Transport; D018341:Receptors, Adrenergic, alpha-2 / Q000032:analysis / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000032:analysis* / Q000378:metabolism*; D015398:Signal Transduction
D018341:Receptors, Adrenergic, alpha-2; D043562:Receptors, G-Protein-Coupled