Beta-arrestin2-mediated internalization of mammalian odorant receptors.
Odorant receptors comprise the biggest subfamily of G-protein-coupled receptors. Although the endocytic mechanisms of other G-protein-coupled receptors have been characterized extensively, almost nothing is known about the intracellular trafficking of odorant receptors. The present study describes the endocytic pathway of mammalian odorant receptors, which bind beta-arrestin2 with high affinity and are internalized via a clathrin-dependent mechanism. After prolonged odorant exposure, receptors are not targeted to lysosomal degradation but accumulate in recycling endosomes. Odorant-induced odorant receptor desensitization is promoted by cAMP-dependent protein kinase A phosphorylation and is dependent on serine and threonine residues within the third intracellular loop of the receptor. Moreover, beta-arrestin2 is redistributed into the dendritic knobs of mouse olfactory receptor neurons after treatment with a complex odorant mixture. Prolonged odorant exposure resulted in accumulation of beta-arrestin2 in intracellular vesicles. Adaptation of olfactory receptor neurons to odorants can be abolished by the inhibition of clathrin-mediated endocytosis, showing the physiological relevance of the here described mechanism of odorant receptor desensitization. A better understanding of odorant receptor trafficking and additional insight into the molecular determinants underlying the interactions of odorant receptors with beta-arrestin2 and other trafficking proteins will therefore be important to fully understand the mechanisms of adaptation and sensitization in the olfactory epithelium.
Mashukova
Anastasia
A
Cell Physiology, Ruhr-Universitaet Bochum, 44780 Bochum, Germany.
Spehr
Marc
M
Hatt
Hanns
H
Neuhaus
Eva M
EM
D020816:Amino Acid Motifs; D000818:Animals; D019390:Arrestins / Q000235:genetics / Q000502:physiology*; D002087:Butyrates / Q000494:pharmacology*; D020013:Calcium Signaling; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D015536:Down-Regulation; D004705:Endocytosis / Q000502:physiology*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism*; D017930:Genes, Reporter; D006185:Habituation, Psychophysiologic; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology; D007668:Kidney / Q000166:cytology / Q000196:embryology; D008024:Ligands; D008297:Male; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009812:Odorants; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D018035:Receptors, Odorant / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D014162:Transfection; D014168:Transferrin / Q000378:metabolism; D000071557:beta-Arrestins
D019390:Arrestins; D002087:Butyrates; D007546:Isoquinolines; D008024:Ligands; C514887:OR2AG1 protein, human; D018035:Receptors, Odorant; D011993:Recombinant Fusion Proteins; D013449:Sulfonamides; D014168:Transferrin; D000071557:beta-Arrestins; C046390:n-amyl butyrate; D017868:Cyclic AMP-Dependent Protein Kinases; D019204:GTP-Binding Proteins; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide