![[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}
|
|
|
|
|
||
CD Wang, MA Buck and CM Fraser
Section on Molecular Neurobiology, National Institute on Alcohol Abuse and Alcoholism, Rockville, Maryland 20852.
Molecular cloning of the alpha 2A-adrenergic receptor has shown that this receptor is a member of the gene superfamily of guanine nucleotide- binding protein (G protein)-coupled receptors. The alpha 2A-adrenergic receptor expressed in Chinese hamster ovary (CHO) cells attenuates and potentiates forskolin-stimulated cAMP production through independent signaling pathways. To examine the role of three conserved aspartic acid and two conserved serine residues in alpha 2A-adrenergic receptor function, we substituted asparagine for aspartic acid or alanine for serine and characterized the mutant receptors in stably transfected CHO cells. Asn113 mutant alpha 2-adrenergic receptors display no [3H] yohimbine specific binding, at concentrations up to 1000 nM. In transfected cells expressing the Asn113 mutant receptor, agonists, at concentrations up to 0.1 mM, produce small decreases (approximately 10% of wild-type values) in forskolin-stimulated cAMP and potentiate forskolin-stimulated cAMP concentrations in a dose-dependent manner, with EC50 values approximately 500-fold higher than those for the wild- type receptor. These findings suggest that Asp113 may be involved in high affinity binding of agonists and antagonists, as has been previously reported for beta 2-adrenergic and m1 muscarinic acetylcholine receptors. Asn79 mutant alpha 2-adrenergic receptors display high affinity agonist binding that is insensitive to guanine nucleotides, suggesting an altered receptor-G protein coupling. Furthermore, agonist binding to Asn79 mutant receptors elicits no change in forskolin-stimulated cAMP concentrations, similar to earlier findings that the corresponding residue in beta 2-adrenergic and muscarinic receptors is required for effector stimulation. Asp130 appears to influence receptor-G protein coupling. Mutation of this residue eliminates high affinity, guanine nucleotide-sensitive, agonist binding and produces a rightward shift in the dose-response curves for agonist-mediated inhibition of forskolin-stimulated cAMP production, compared with the wild-type receptor. Moreover, agonist potentiation of forskolin-stimulated cAMP levels is abolished if Asp130 is replaced by Asn, supporting the hypothesis that inhibition and potentiation of forskolin-stimulated cAMP production in CHO cells proceed through distinct signaling pathways. Characterization of Ala204 mutant alpha 2A- adrenergic receptors suggests a possible role for Ser204 in hydrogen bond interactions with the para-hydroxyl group of the phenyl ring of the catecholamines, as has been previously described for the corresponding serine in beta 2-adrenergic receptors.(ABSTRACT TRUNCATED AT 400 WORDS)
This article has been cited by other articles:
|
|
 |
|
  M. R. Braden and D. E. Nichols Assessment of the Roles of Serines 5.43(239) and 5.46(242) for Binding and Potency of Agonist Ligands at the Human Serotonin 5-HT2A Receptor Mol. Pharmacol., November 1, 2007; 72(5): 1200 - 1209. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. E. Rovati, V. Capra, and R. R. Neubig The Highly Conserved DRY Motif of Class A G Protein-Coupled Receptors: Beyond the Ground State Mol. Pharmacol., April 1, 2007; 71(4): 959 - 964. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. K. Yee, A. Suzuki, L. Luo, and S. J. Fluharty Identification of Structural Determinants for G Protein-Independent Activation of Mitogen-Activated Protein Kinases in the Seventh Transmembrane Domain of the Angiotensin II Type 1 Receptor Mol. Endocrinol., August 1, 2006; 20(8): 1924 - 1934. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. T. Ukairo, C. D. Bondi, A. H. Newman, S. S. Kulkarni, A. P. Kozikowski, S. Pan, and C. K. Surratt Recognition of Benztropine by the Dopamine Transporter (DAT) Differs from That of the Classical Dopamine Uptake Inhibitors Cocaine, Methylphenidate, and Mazindol as a Function of a DAT Transmembrane 1 Aspartic Acid Residue J. Pharmacol. Exp. Ther., August 1, 2005; 314(2): 575 - 583. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Li, N. M. Nowak, S.-K. Kim, K. A. Jacobson, A. Bagheri, C. Schmidt, and J. Wess Random Mutagenesis of the M3 Muscarinic Acetylcholine Receptor Expressed in Yeast: IDENTIFICATION OF SECOND-SITE MUTATIONS THAT RESTORE FUNCTION TO A COUPLING-DEFICIENT MUTANT M3 RECEPTOR J. Biol. Chem., February 18, 2005; 280(7): 5664 - 5675. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. P. Millar, Z.-L. Lu, A. J. Pawson, C. A. Flanagan, K. Morgan, and S. R. Maudsley Gonadotropin-Releasing Hormone Receptors Endocr. Rev., April 1, 2004; 25(2): 235 - 275. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Schumann, T. Nakagawa, S. A. Mantey, K. Tokita, D. J. Venzon, S. J. Hocart, R. V. Benya, and R. T. Jensen Importance of Amino Acids of the Central Portion of the Second Intracellular Loop of the Gastrin-Releasing Peptide Receptor for Phospholipase C Activation, Internalization, and Chronic Down-Regulation J. Pharmacol. Exp. Ther., November 1, 2003; 307(2): 597 - 607. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. S. Huang Construction of a sequence motif characteristic of aminergic G protein-coupled receptors Protein Sci., July 1, 2003; 12(7): 1360 - 1367. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. J. Pauwels, I. Rauly, and T. Wurch Dissimilar Pharmacological Responses by a New Series of Imidazoline Derivatives at Precoupled and Ligand-Activated {alpha}2A-Adrenoceptor States: Evidence for Effector Pathway-Dependent Differential Antagonism J. Pharmacol. Exp. Ther., June 1, 2003; 305(3): 1015 - 1023. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Shin, E. Coates, N. J. Murgolo, K. L. Morse, M. Bayne, C. D. Strader, and F. J. Monsma Jr. Molecular Modeling and Site-Specific Mutagenesis of the Histamine-Binding Site of the Histamine H4 Receptor Mol. Pharmacol., July 1, 2002; 62(1): 38 - 47. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. J. Uveges, D. Kowal, Y. Zhang, T. B. Spangler, J. Dunlop, S. Semus, and P. G. Jones The Role of Transmembrane Helix 5 in Agonist Binding to the Human H3 Receptor J. Pharmacol. Exp. Ther., May 1, 2002; 301(2): 451 - 458. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. S. Nelson, M. Ikeda, H. S. Gompf, M. L. Robinson, N. K. Fuchs, T. Yoshioka, K. A. Neve, and C. N. Allen Regulation of Melatonin 1a Receptor Signaling and Trafficking by Asparagine-124 Mol. Endocrinol., August 1, 2001; 15(8): 1306 - 1317. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Ballesteros, L. Shi, and J. A. Javitch Structural Mimicry in G Protein-Coupled Receptors: Implications of the High-Resolution Structure of Rhodopsin for Structure-Function Analysis of Rhodopsin-Like Receptors Mol. Pharmacol., July 1, 2001; 60(1): 1 - 19. [Abstract] [Full Text]
|
|
|
|
|
|
T. Nyrönen, M. Pihlavisto, J. M. Peltonen, A.-M. Hoffrén, M. Varis, T. Salminen, S. Wurster, A. Marjamäki, L. Kanerva, E. Katainen, et al. Molecular Mechanism for Agonist-Promoted alpha 2A-Adrenoceptor Activation by Norepinephrine and Epinephrine Mol. Pharmacol., April 16, 2001; 59(5): 1343 - 1354. [Abstract] [Full Text]
|
|
|
|
 |
|
 S. A. Oliveira and T. E. Shenk Murine cytomegalovirus M78 protein, a G protein-coupled receptor homologue, is a constituent of the virion and facilitates accumulation of immediate-early viral mRNA PNAS, March 13, 2001; 98(6): 3237 - 3242. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. M. Topham, L. Mouledous, and J.-C. Meunier On the spatial disposition of the fifth transmembrane helix and the structural integrity of the transmembrane binding site in the opioid and ORL1 G protein-coupled receptor family Protein Eng. Des. Sel., July 1, 2000; 13(7): 477 - 490. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Allman, K. M. Page, C. A.M. Curtis, and E. C. Hulme Scanning Mutagenesis Identifies Amino Acid Side Chains in Transmembrane Domain 5 of the M1 Muscarinic Receptor that Participate in Binding the Acetyl Methyl Group of Acetylcholine Mol. Pharmacol., July 1, 2000; 58(1): 175 - 184. [Abstract] [Full Text]
|
|
|
|
|
|
D. Macdonald, N. Murgolo, R. Zhang, J. P. Durkin, X. Yao, C. D. Strader, and M. P. Graziano Molecular Characterization of the Melanin-Concentrating Hormone/Receptor Complex: Identification of Critical Residues Involved in Binding and Activation Mol. Pharmacol., July 1, 2000; 58(1): 217 - 225. [Abstract] [Full Text]
|
|
|
|
|
|
J. E. Porter and D. M. Perez Influence of a Lysine 331 Counterion on the pKa of Aspartic Acid 125: Evidence for a Salt-Bridge Interaction and Role in alpha 1b-Adrenergic Receptor Activation J. Pharmacol. Exp. Ther., January 1, 2000; 292(1): 440 - 448. [Abstract] [Full Text]
|
|
|
|
|
|
C. Ambrosio, P. Molinari, S. Cotecchia, and T. Costa Catechol-Binding Serines of beta 2-Adrenergic Receptors Control the Equilibrium between Active and Inactive Receptor States Mol. Pharmacol., January 1, 2000; 57(1): 198 - 210. [Abstract] [Full Text]
|
|
|
|
|
|
J. E. Porter and D. M. Perez Characteristics for a Salt-bridge Switch Mutation of the alpha 1b Adrenergic Receptor. ALTERED PHARMACOLOGY AND RESCUE OF CONSTITUTIVE ACTIVITY J. Biol. Chem., December 3, 1999; 274(49): 34535 - 34538. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Roche, S. Bounds, S. Brown, and K. Mackie A Mutation in the Second Transmembrane Region of the CB1 Receptor Selectively Disrupts G Protein Signaling and Prevents Receptor Internalization Mol. Pharmacol., September 1, 1999; 56(3): 611 - 618. [Abstract] [Full Text]
|
|
|
|
|
|
T. Salminen, M. Varis, T. Nyronen, M. Pihlavisto, A.-M. Hoffren, T. Lonnberg, A. Marjamaki, H. Frang, J.-M. Savola, M. Scheinin, et al. Three-dimensional Models of alpha 2A-Adrenergic Receptor Complexes Provide a Structural Explanation for Ligand Binding J. Biol. Chem., August 13, 1999; 274(33): 23405 - 23413. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. J. Slotboom, W. N. Konings, and J. S. Lolkema Structural Features of the Glutamate Transporter Family Microbiol. Mol. Biol. Rev., June 1, 1999; 63(2): 293 - 307. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. J. Doranz, M. J. Orsini, J. D. Turner, T. L. Hoffman, J. F. Berson, J. A. Hoxie, S. C. Peiper, L. F. Brass, and R. W. Doms Identification of CXCR4 Domains That Support Coreceptor and Chemokine Receptor Functions J. Virol., April 1, 1999; 73(4): 2752 - 2761. [Abstract] [Full Text]
|
|
|
|
|
|
C. Saunders, J. R. Keefer, C. A. Bonner, and L. E. Limbird Targeting of G Protein-coupled Receptors to the Basolateral Surface of Polarized Renal Epithelial Cells Involves Multiple, Non-contiguous Structural Signals J. Biol. Chem., September 11, 1998; 273(37): 24196 - 24206. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. L. Wiens, C. S. Nelson, and K. A. Neve Contribution of Serine Residues to Constitutive and Agonist-Induced Signaling via the D2S Dopamine Receptor: Evidence for Multiple, Agonist-Specific Active Conformations Mol. Pharmacol., August 1, 1998; 54(2): 435 - 444. [Abstract] [Full Text]
|
|
|
|
|
|
A. Marjamäki, M. Pihlavisto, V. Cockcroft, P. Heinonen, J.-M. Savola, and M. Scheinin Chloroethylclonidine Binds Irreversibly to Exposed Cysteines in the Fifth Membrane-Spanning Domain of the Human alpha 2A-Adrenergic Receptor Mol. Pharmacol., March 1, 1998; 53(3): 370 - 376. [Abstract] [Full Text]
|
|
|
|
 |
|
 C. MISSALE, S. R. NASH, S. W. ROBINSON, M. JABER, and M. G. CARON Dopamine Receptors: From Structure to Function Physiol Rev, January 1, 1998; 78(1): 189 - 225. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. E. Spivak, C. L. Beglan, B. K. Seidleck, L. D. Hirshbein, C. J. Blaschak, G. R. Uhl, and C. K. Surratt Naloxone Activation of µ-Opioid Receptors Mutated at a Histidine Residue Lining the Opioid Binding Cavity Mol. Pharmacol., December 1, 1997; 52(6): 983 - 992. [Abstract] [Full Text]
|
|
|
|
|
|
F. G. Boess, F. J. Monsma Jr., V. Meyer, C. Zwingelstein, and A. J. Sleight Interaction of Tryptamine and Ergoline Compounds with Threonine 196 in the Ligand Binding Site of the 5-Hydroxytryptamine6 Receptor Mol. Pharmacol., September 1, 1997; 52(3): 515 - 523. [Abstract] [Full Text]
|
|
|
|
|
|
K. K. Arora, Z. Cheng, and K. J. Catt Mutations of the Conserved DRS Motif in the Second Intracellular Loop of the Gonadotropin-Releasing Hormone Receptor Affect Expression, Activation, and Internalization Mol. Endocrinol., August 1, 1997; 11(9): 1203 - 1212. [Abstract] [Full Text]
|
|
|
|
|
|
S. C. Sealfon, H. Weinstein, and R. P. Millar Molecular Mechanisms of Ligand Interaction with the Gonadotropin-Releasing Hormone Receptor Endocr. Rev., April 1, 1997; 18(2): 180 - 205. [Abstract] [Full Text]
|
|
|
|
|
|
C. C. Gerhardt, H. C. Lodder, M. Vincent, R. A. Bakker, R. J. Planta, E. Vreugdenhil, K. S. Kits, and H. van Heerikhuizen Cloning and Expression of a Complementary DNA Encoding a Molluscan Octopamine Receptor That Couples to Chloride Channels in HEK293 Cells J. Biol. Chem., March 7, 1997; 272(10): 6201 - 6207. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-L. Lu, C. A. Curtis, P. G. Jones, J. Pavia, and E. C. Hulme The Role of the Aspartate-Arginine-Tyrosine Triad in the m1 Muscarinic Receptor: Mutations of Aspartate 122 and Tyrosine 124 Decrease Receptor Expression but Do Not Abolish Signaling Mol. Pharmacol., February 1, 1997; 51(2): 234 - 241. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. C. Gerhardt, R. A. Bakker, G. J. Piek, R. J. Planta, E. Vreugdenhil, J. E. Leysen, and H. Van Heerikhuizen Molecular Cloning and Pharmacological Characterization of a Molluscan Octopamine Receptor Mol. Pharmacol., February 1, 1997; 51(2): 293 - 300. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. E. Porter, J. Hwa, and D. M. Perez Activation of the alpha 1b-Adrenergic Receptor Is Initiated by Disruption of an Interhelical Salt Bridge Constraint J. Biol. Chem., November 8, 1996; 271(45): 28318 - 28323. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Acharya and S. S. Karnik Modulation of GDP Release from Transducin by the Conserved Glu134-Arg135 Sequence in Rhodopsin J. Biol. Chem., October 11, 1996; 271(41): 25406 - 25411. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Almaula, B. J. Ebersole, D. Zhang, H. Weinstein, and S. C. Sealfon Mapping the Binding Site Pocket of the Serotonin 5-Hydroxytryptamine2A Receptor. Ser3.36(159) PROVIDES A SECOND INTERACTION SITE FOR THE PROTONATED AMINE OF SEROTONIN BUT NOT OF LYSERGIC ACID DIETHYLAMIDE OR BUFOTENIN J. Biol. Chem., June 21, 1996; 271(25): 14672 - 14675. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Hwa and D. M. Perez The Unique Nature of the Serine Interactions for alpha(1)-Adrenergic Receptor Agonist Binding and Activation J. Biol. Chem., March 15, 1996; 271(11): 6322 - 6327. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. K. Arora, A. Sakai, and K. J. Catt Effects of Second Intracellular Loop Mutations on Signal Transduction and Internalization of the Gonadotropin-releasing Hormone Receptor J. Biol. Chem., September 29, 1995; 270(39): 22820 - 22826. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Hwa, R. M. Graham, and D. M. Perez Identification of Critical Determinants of alpha(1)-Adrenergic Receptor Subtype Selective Agonist Binding J. Biol. Chem., September 29, 1995; 270(39): 23189 - 23195. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. C. Sealfon, L. Chi, B. J. Ebersole, V. Rodic, D. Zhang, J. A. Ballesteros, and H. Weinstein Related Contribution of Specific Helix 2 and 7 Residues to Conformational Activation of the Serotonin 5-HT[IMAGE] Receptor J. Biol. Chem., July 14, 1995; 270(28): 16683 - 16688. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A Surprenant, D. Horstman, H Akbarali, and L. Limbird A point mutation of the alpha 2-adrenoceptor that blocks coupling to potassium but not calcium currents Science, August 14, 1992; 257(5072): 977 - 980. [Abstract] [PDF]
|
|
|
|
|
|
G. Liapakis, J. A. Ballesteros, S. Papachristou, W. C. Chan, X. Chen, and J. A. Javitch The Forgotten Serine. A CRITICAL ROLE FOR Ser-2035.42 IN LIGAND BINDING TO AND ACTIVATION OF THE beta 2-ADRENERGIC RECEPTOR J. Biol. Chem., November 22, 2000; 275(48): 37779 - 37788. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Frang, V. Cockcroft, T. Karskela, M. Scheinin, and A. Marjamaki Phenoxybenzamine Binding Reveals the Helical Orientation of the Third Transmembrane Domain of Adrenergic Receptors J. Biol. Chem., August 10, 2001; 276(33): 31279 - 31284. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
