Agonist-dependent dissociation of human somatostatin receptor 2 dimers: a role in receptor trafficking.
G-protein-coupled receptors (GPCRs) represent the largest and most diverse family of cell surface receptors. Several GPCRs have been documented to dimerize with resulting changes in pharmacology and signaling. We have previously reported, by means of photobleaching fluorescence resonance energy transfer (pbFRET) microscopy and fluorescence correlation spectroscopic analysis in live cells, that human somatostatin receptor (hSSTR) 5 could both homodimerize and heterodimerize with hSSTR1 in the presence of the agonist SST-14. By contrast, hSSTR1 remained monomeric when expressed alone regardless of agonist exposure in live cells. However, the effect of the agonist on other hSSTR members remains unknown. Using pbFRET microscopy and Western blot, we provide evidence for agonist-dependent dissociation of self-associated hSSTR2 stably expressed in CHO-K1 and HEK-293 cells. Furthermore, the dissociation of the hSSTR2 dimer occurred in a concentration-dependent manner. Moreover, blocking receptor dissociation using a cross-linker agent perturbed receptor trafficking. Taking these data together, we suggest that the process of GPCR dimerization may operate differently, even among members of the same family, and that receptor dissociation as well as dimerization may be important steps for receptor dynamics.
Grant
Michael
M
Fraser Laboratories for Diabetes Research, Department of Medicine, Royal Victoria Hospital, 687 Pine Avenue West, Montreal, Quebec H3A 1A1, Canada.
Collier
Brian
B
Kumar
Ujendra
U
D000818:Animals; D015153:Blotting, Western; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000494:pharmacology; D019281:Dimerization; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007150:Immunohistochemistry; D007700:Kinetics; D008024:Ligands; D010455:Peptides / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry; D017481:Receptors, Somatostatin / Q000819:agonists* / Q000737:chemistry*; D013053:Spectrophotometry; D013997:Time Factors
D003432:Cross-Linking Reagents; D008024:Ligands; D010455:Peptides; D043562:Receptors, G-Protein-Coupled; D017481:Receptors, Somatostatin; C101130:somatostatin receptor type 1; C093733:somatostatin receptor 2