MolPharm xPharm- The Comprehensive Pharmacology Reference

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Roche, J. P.
Right arrow Articles by Mackie, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Roche, J. P.
Right arrow Articles by Mackie, K.

Vol. 56, Issue 3, 611-618, September 1999

A Mutation in the Second Transmembrane Region of the CB1 Receptor Selectively Disrupts G Protein Signaling and Prevents Receptor Internalization

John P. Roche, Sid Bounds, Sean Brown, and Ken Mackie

Departments of Anesthesiology (Se.B., K.M.), and Physiology and Biophysics (J.P.R., K.M.), University of Washington School of Medicine, Seattle, Washington; and MDS Panlabs, Bothell, Washington (Si.B.)

We mutated a conserved aspartate in the second transmembrane domain of the cannabinoid CB1 receptor to asparagine (D164N), stably transfected it into AtT20 cells, and examined the coupling of this mutant receptor to several intracellular effectors that are targets of wild-type CB1 receptor activation. We found that the D164N receptor binds the CB1 agonist WIN 55,212-2 with an affinity matching that of the wild-type CB1 receptor and inhibits Ca2+ currents and cAMP production with an equivalent potency and efficacy. This mutation, however, blocks coupling of the receptor to the potentiation of inwardly rectifying potassium channel (KIR) currents and prevents internalization of the receptor after exposure to agonist. Although the mutant receptor did not internalize, we found it was still capable of activating p42/44 MAP kinase. In addition, we made a reciprocal mutation that exchanged the aspartate with an asparagine in the seventh transmembrane region (D164N/N394D). In other seven-membrane-spanning receptors, this reciprocal mutation is known to restore functions disrupted by the mutation of the single conserved aspartate. However, activation of D164N/N394D did not potentiate KIR current, nor did it internalize. We conclude that D164 is necessary for potentiation of KIR current and internalization of receptor but not necessary for agonist binding, inhibition of cAMP production, inhibition of Ca2+ currents, or activation of p42/44 MAP kinase. Furthermore, CB1 receptor internalization is not necessary for MAP kinase activation.

Copyright © 1999 by The American Society for Pharmacology and Experimental Therapeutics


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
M. Canals and G. Milligan
Constitutive Activity of the Cannabinoid CB1 Receptor Regulates the Function of Co-expressed Mu Opioid Receptors
J. Biol. Chem., April 25, 2008; 283(17): 11424 - 11434.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
A. Kapur, P. Samaniego, G. A. Thakur, A. Makriyannis, and M. E. Abood
Mapping the Structural Requirements in the CB1 Cannabinoid Receptor Transmembrane Helix II for Signal Transduction
J. Pharmacol. Exp. Ther., April 1, 2008; 325(1): 341 - 348.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Anavi-Goffer, D. Fleischer, D. P. Hurst, D. L. Lynch, J. Barnett-Norris, S. Shi, D. L. Lewis, S. Mukhopadhyay, A. C. Howlett, P. H. Reggio, et al.
Helix 8 Leu in the CB1 Cannabinoid Receptor Contributes to Selective Signal Transduction Mechanisms
J. Biol. Chem., August 24, 2007; 282(34): 25100 - 25113.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
N. A. McDonald, C. M. Henstridge, C. N. Connolly, and A. J. Irving
An Essential Role for Constitutive Endocytosis, but Not Activity, in the Axonal Targeting of the CB1 Cannabinoid Receptor
Mol. Pharmacol., April 1, 2007; 71(4): 976 - 984.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Leterrier, D. Bonnard, D. Carrel, J. Rossier, and Z. Lenkei
Constitutive Endocytic Cycle of the CB1 Cannabinoid Receptor
J. Biol. Chem., August 20, 2004; 279(34): 36013 - 36021.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
I. Galve-Roperh, D. Rueda, T. Gomez del Pulgar, G. Velasco, and M. Guzman
Mechanism of Extracellular Signal-Regulated Kinase Activation by the CB1 Cannabinoid Receptor
Mol. Pharmacol., December 1, 2002; 62(6): 1385 - 1392.
[Abstract] [Full Text] [PDF]

Home page
 Pharmacol. Rev.Home page
A. C. Howlett, F. Barth, T. I. Bonner, G. Cabral, P. Casellas, W. A. Devane, C. C. Felder, M. Herkenham, K. Mackie, B. R. Martin, et al.
International Union of Pharmacology. XXVII. Classification of Cannabinoid Receptors
Pharmacol. Rev., June 1, 2002; 54(2): 161 - 202.
[Abstract] [Full Text] [PDF]

Home page
 FASEB J.Home page
R. RIMONDINI, C. ARLINDE, W. SOMMER, and M. HEILIG
Long-lasting increase in voluntary ethanol consumption and transcriptional regulation in the rat brain after intermittent exposure to alcohol
FASEB J, January 1, 2002; 16(1): 27 - 35.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
J. Nie and D. L. Lewis
Structural Domains of the CB1 Cannabinoid Receptor That Contribute to Constitutive Activity and G-Protein Sequestration
J. Neurosci., November 15, 2001; 21(22): 8758 - 8764.
[Abstract] [Full Text] [PDF]

Home page
 J. Neurosci.Home page
A. A. Coutts, S. Anavi-Goffer, R. A. Ross, D. J. MacEwan, K. Mackie, R. G. Pertwee, and A. J. Irving
Agonist-Induced Internalization and Trafficking of Cannabinoid CB1 Receptors in Hippocampal Neurons
J. Neurosci., April 1, 2001; 21(7): 2425 - 2433.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
M. H. Wilson, H. A. Highfield, and L. E. Limbird
The Role of a Conserved Inter-Transmembrane Domain Interface in Regulating {alpha}2a-Adrenergic Receptor Conformational Stability and Cell-Surface Turnover
Mol. Pharmacol., April 1, 2001; 59(4): 929 - 938.
[Abstract] [Full Text]

Home page
 Proc. Natl. Acad. Sci. USAHome page
K. L. Pierce, S. Maudsley, Y. Daaka, L. M. Luttrell, and R. J. Lefkowitz
Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors
PNAS, February 15, 2000; 97(4): 1489 - 1494.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 1999 by the American Society for Pharmacology and Experimental Therapeutics