Constitutively active GPR6 is located in the intracellular compartments

We used multiple imaging assays to test the hypothesis that GPR6, a constitutively active Gs-coupled receptor, is present on the cell surface. A pHluorin tag at the N-terminus of rat GPR6 expressed in human embryonic kidney 293 (HEK293) cells was not accessible to protons, chymotrypsin or anti-green...

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Bibliographic Details
Published in:FEBS letters 2009-01, Vol.583 (1), p.107-112
Main Authors: Padmanabhan, Shalini, Myers, Amanda G., Prasad, Balakrishna M.
Format: Article
Language:English
Subjects:
Animals
cAMP
cAMP response element-binding protein
Cell Line
Cell Membrane
Constitutive activity
Corpus Striatum - cytology
Corpus Striatum - metabolism
CREB
cyclic adenosine monophosphate
Cyclic AMP
G-protein coupled receptor
GFP
GPCR
green fluorescent protein
HEK293
human embryonic kidney 293
Humans
Neuron
Neurons - metabolism
Plasma membrane
Rats
Receptors, G-Protein-Coupled - genetics
Receptors, G-Protein-Coupled - metabolism
Striatum
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Summary:We used multiple imaging assays to test the hypothesis that GPR6, a constitutively active Gs-coupled receptor, is present on the cell surface. A pHluorin tag at the N-terminus of rat GPR6 expressed in human embryonic kidney 293 (HEK293) cells was not accessible to protons, chymotrypsin or anti-green fluorescent protein antibody, demonstrating that GPR6 is primarily located in intracellular compartments. Similar intracellular localization of pHluorin-tagged GPR6 was found in striatal neurons, where endogenous GPR6 is expressed. Confirmation of Gs-mediated constitutive activity in HEK293 cells and striatal neurons led us to conclude that GPR6 can signal from intracellular compartments.
ISSN:0014-5793
1873-3468
DOI:10.1016/j.febslet.2008.11.033