![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 56, Issue 5, 1005-1013, November 1999
2A-Adrenergic
Receptors: Distinct Basic Residues Mediate Gi versus
Gs Activation
Departments of Pharmacology (S.M.W, W.K.L, K.-L.L, D.A.C, M.N.,
R.R.N.) and Internal Medicine/Hypertension (R.R.N.), Biophysics
Research Division (D.A.C, R.R.N.), The University of Michigan, Ann
Arbor, Michigan
The structural determinants of G protein coupling versus activation by
G protein-coupled receptors are not well understood. We examine the
role of two distinct basic regions in the carboxyl terminal portion of
the third intracellular loop of the 
2A-adrenergic receptor to dissect these aspects of function. Changing three arginines
to alanines by mutagenesis and stable expression in Chinese hamster
ovary-K1 cells impaired the 2-adrenergic receptor Gs-mediated stimulation of cyclic AMP (cAMP) accumulation,
whereas Gi-mediated inhibition was normal. When two (B2) or
three (B3) basic residues closer to transmembrane span 6 were mutated
to alanine, normal ligand binding was observed, but
Gi-mediated inhibition of cAMP accumulation showed 20-fold
and 50-fold decreases in agonist potency for the B2 and B3 mutants,
respectively. Surprisingly, a normal Gs response was seen
for the B2 mutant, and the B3 mutant showed only a 6-fold decrease in
agonist potency. Mutation of both the three alanines and B3 residues to
alanines showed a 200-fold decrease in agonist potency for
Gi-mediated inhibition of cAMP accumulation, whereas the
Gs response was nearly completely eliminated. The three
basic residues (which include the BB of the BBXXF motif) play a role as
Gi activators rather than in receptor-G protein coupling,
because high-affinity agonist binding is intact. Thus, we have
identified three basic residues required for activation of
Gi but not required for receptor-G protein coupling. Also, distinct basic residues are required for optimal Gi and
Gs responses, defining a microspecificity determinant
within the carboxyl terminal portion of the third intracellular loop of
the 2a adrenergic receptor.
This article has been cited by other articles:
|
|
 |
|
  G. S. Lynch and J. G. Ryall Role of {beta}-Adrenoceptor Signaling in Skeletal Muscle: Implications for Muscle Wasting and Disease Physiol Rev, April 1, 2008; 88(2): 729 - 767. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Huang, Y. Fu, R. A. Charbeneau, T. L. Saunders, D. K. Taylor, K. D. Hankenson, M. W. Russell, L. G. D'Alecy, and R. R. Neubig Pleiotropic Phenotype of a Genomic Knock-In of an RGS-Insensitive G184S Gnai2 Allele. Mol. Cell. Biol., September 1, 2006; 26(18): 6870 - 6879. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Muramatsu and M. Suwa Statistical analysis and prediction of functional residues effective for GPCR-G-protein coupling selectivity Protein Eng. Des. Sel., June 1, 2006; 19(6): 277 - 283. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Maudsley, B. Martin, and L. M. Luttrell The Origins of Diversity and Specificity in G Protein-Coupled Receptor Signaling J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 485 - 494. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Bodenstein, D. P. Venter, and C. B. Brink Phenoxybenzamine and Benextramine, but Not 4-Diphenylacetoxy-N-[2-chloroethyl]piperidine Hydrochloride, Display Irreversible Noncompetitive Antagonism at G Protein-Coupled Receptors J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 891 - 905. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Saito, M. Tetsuka, S. Saito, K. Imai, A. Yoshikawa, H. Doi, and K. Maruyama Arginine Residue 155 in the Second Intracellular Loop Plays a Critical Role in Rat Melanin-Concentrating Hormone Receptor 1 Activation Endocrinology, August 1, 2005; 146(8): 3452 - 3462. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Tetsuka, Y. Saito, K. Imai, H. Doi, and K. Maruyama The Basic Residues in the Membrane-Proximal C-Terminal Tail of the Rat Melanin-Concentrating Hormone Receptor 1 Are Required for Receptor Function Endocrinology, August 1, 2004; 145(8): 3712 - 3723. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Lee, A. Robeva, Z. Chen, and K. P. Minneman Mutational Uncoupling of {alpha}1A-Adrenergic Receptors from G Proteins Also Uncouples Mitogenic and Transcriptional Responses in PC12 Cells J. Pharmacol. Exp. Ther., August 1, 2003; 306(2): 471 - 477. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. M. Berglund, D. A. Schober, M. A. Statnick, P. H. McDonald, and D. R. Gehlert The Use of Bioluminescence Resonance Energy Transfer 2 to Study Neuropeptide Y Receptor Agonist-Induced {beta}-Arrestin 2 Interaction J. Pharmacol. Exp. Ther., July 1, 2003; 306(1): 147 - 156. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C.-Y. Lin, M. G. Varma, A. Joubel, S. Madabushi, O. Lichtarge, and D. L. Barber Conserved Motifs in Somatostatin, D2-dopamine, and alpha 2B-Adrenergic Receptors for Inhibiting the Na-H Exchanger, NHE1 J. Biol. Chem., April 18, 2003; 278(17): 15128 - 15135. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Zhong, S. M. Wade, P. J. Woolf, J. J. Linderman, J. R. Traynor, and R. R. Neubig A Spatial Focusing Model for G Protein Signals. REGULATOR OF G PROTEIN SIGNALING (RGS) PROTEIN-MEDIATED KINETIC SCAFFOLDING J. Biol. Chem., February 21, 2003; 278(9): 7278 - 7284. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. DeGraff, V. V. Gurevich, and J. L. Benovic The Third Intracellular Loop of alpha 2-Adrenergic Receptors Determines Subtype Specificity of Arrestin Interaction J. Biol. Chem., November 1, 2002; 277(45): 43247 - 43252. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Christopoulos and T. Kenakin G Protein-Coupled Receptor Allosterism and Complexing Pharmacol. Rev., June 1, 2002; 54(2): 323 - 374. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. H. Berlot A Highly Effective Dominant Negative alpha s Construct Containing Mutations That Affect Distinct Functions Inhibits Multiple Gs-coupled Receptor Signaling Pathways J. Biol. Chem., May 31, 2002; 277(23): 21080 - 21085. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. J. Miller, X. Shi, and H. Burkin Molecular Basis of Mammalian Gamete Binding Recent Prog. Horm. Res., January 1, 2002; 57(1): 37 - 73. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. C. Jeyaraj, M. A. Chotani, S. Mitra, H. E. Gregg, N. A. Flavahan, and K. J. Morrison Cooling Evokes Redistribution of alpha 2C-Adrenoceptors from Golgi to Plasma Membrane in Transfected Human Embryonic Kidney 293 Cells Mol. Pharmacol., December 1, 2001; 60(6): 1195 - 1200. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. G. Modrall, M. Nanamori, J. Sadoshima, D. C. Barnhart, J. C. Stanley, and R. R. Neubig ANG II type 1 receptor downregulation does not require receptor endocytosis or G protein coupling Am J Physiol Cell Physiol, September 1, 2001; 281(3): C801 - C809. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Watson, G. Chen, P. Irving, J. Way, W.-J. Chen, and T. Kenakin The Use of Stimulus-Biased Assay Systems to Detect Agonist-Specific Receptor Active States: Implications for the Trafficking of Receptor Stimulus by Agonists Mol. Pharmacol., April 13, 2001; 58(6): 1230 - 1238. [Abstract] [Full Text]
|
|
|
|
 |
|
 T. KENAKIN Inverse, protean, and ligand-selective agonism: matters of receptor conformation FASEB J, March 1, 2001; 15(3): 598 - 611. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Wade, K.-L. Lan, D. J. Moore, and R. R. Neubig Inverse Agonist Activity at the {alpha}2A-Adrenergic Receptor Mol. Pharmacol., March 1, 2001; 59(3): 532 - 542. [Abstract] [Full Text]
|
|
|
|
 |
|
 X Shi, S Amindari, K Paruchuru, D Skalla, H Burkin, B. Shur, and D. Miller Cell surface beta-1,4-galactosyltransferase-I activates G protein-dependent exocytotic signaling Development, January 3, 2001; 128(5): 645 - 654. [Abstract] [PDF]
|
|
|
|
|
|
M. J. Millan, A. Dekeyne, A. Newman-Tancredi, D. Cussac, V. Audinot, G. Milligan, D. Duqueyroix, S. Girardon, J. Mullot, J. A. Boutin, et al. S18616, a Highly Potent, Spiroimidazoline Agonist at alpha 2-Adrenoceptors: I. Receptor Profile, Antinociceptive and Hypothermic Actions in Comparison with Dexmedetomidine and Clonidine J. Pharmacol. Exp. Ther., December 1, 2000; 295(3): 1192 - 1205. [Abstract] [Full Text]
|
|
|
|
|
|
F. Buck, W. Wang, S. Harder, C. Brathwaite, T. O. Bruhn, and M. C. Gershengorn Juxtamembrane Regions in the Third Intracellular Loop of the Thyrotropin-Releasing Hormone Receptor Type 1 Are Important for Coupling to Gq Endocrinology, October 1, 2000; 141(10): 3717 - 3722. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. B. Brink, S. M. Wade, and R. R. Neubig Agonist-Directed Trafficking of Porcine alpha 2A-Adrenergic Receptor Signaling in Chinese Hamster Ovary Cells: l-Isoproterenol Selectively Activates Gs J. Pharmacol. Exp. Ther., August 1, 2000; 294(2): 539 - 547. [Abstract] [Full Text]
|
|
|
|
|
|
M. R. Mazzoni, S. Taddei, L. Giusti, P. Rovero, C. Galoppini, A. D'Ursi, S. Albrizio, A. Triolo, E. Novellino, G. Greco, et al. A Galpha s Carboxyl-Terminal Peptide Prevents Gs Activation by the A2A Adenosine Receptor Mol. Pharmacol., July 1, 2000; 58(1): 226 - 236. [Abstract] [Full Text]
|
|
 |
 |
 |
|
 |
 |
 |
