![[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. 54, Issue 3, 485-494, September 1998
Department of Pharmacology and Curriculum in Neurobiology,
University of North Carolina School of Medicine, Chapel Hill, North
Carolina 27599
The Gq/phospholipase C-linked human P2Y2
receptor was tagged at its amino terminus with the hemagglutinin A (HA)
epitope sequence (P2Y2-HA) and stably expressed in 1321N1
human astrocytoma cells. Neither the pharmacological selectivity nor
the signaling properties of the receptor were altered by the presence
of the epitope. An enzyme-linked immunosorbent assay was developed to
quantify cell surface levels of P2Y2-HA receptors using an
anti-HA antibody. Incubation of cells with P2Y2 receptor
agonists resulted in a concentration of agonist- and time-dependent
decrease in cell surface immunoreactivity. Methodology for indirect
immunofluorescence confocal microscopy was developed and applied to
demonstrate that the agonist-promoted decreases in cell surface
immunoreactivity paralleled increases in intracellular
immunoreactivity. Agonist-induced internalization of P2Y2
receptors was demonstrated directly by prelabeling P2Y2-HA
receptors with antibody before agonist challenge and then quantifying
the movement of receptors from a cell surface to intracellular
localization in the presence of agonist. Removal of agonist from the
medium resulted in recovery of cell surface immunoreactivity to control
levels within ~1 hr. Incubation of P2Y2-HA
receptor-expressing cells with P2Y2 receptor agonists also
resulted in receptor-specific desensitization of nucleotide-promoted inositol phosphate accumulation. This loss of responsiveness occurred more rapidly and to a greater extent than did the agonist-promoted loss
of surface receptors. Inhibition of receptor internalization by
reduction of temperature to 16° had no effect on the capacity of
nucleotides to induce P2Y2 receptor-specific
desensitization. These results illustrate that the P2Y2
receptor undergoes agonist-promoted movement to an intracellular
compartment. This receptor internalization is not required for
agonist-induced desensitization.
This article has been cited by other articles:
|
|
 |
|
  C. Hoffmann, N. Ziegler, S. Reiner, C. Krasel, and M. J. Lohse Agonist-selective, Receptor-specific Interaction of Human P2Y Receptors with {beta}-Arrestin-1 and -2 J. Biol. Chem., November 7, 2008; 283(45): 30933 - 30941. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Govindaraju, J. G. Martin, K. Maghni, P. Ferraro, and M.-C. Michoud The Effects of Extracellular Purines and Pyrimidines on Human Airway Smooth Muscle Cells J. Pharmacol. Exp. Ther., November 1, 2005; 315(2): 941 - 948. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-D. Qi, S. C. Wolff, and R. A. Nicholas The Apical Targeting Signal of the P2Y2 Receptor Is Located in Its First Extracellular Loop J. Biol. Chem., August 12, 2005; 280(32): 29169 - 29175. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Weick, P. Wiedemann, A. Reichenbach, and A. Bringmann Resensitization of P2Y Receptors by Growth Factor-Mediated Activation of the Phosphatidylinositol-3 Kinase in Retinal Glial Cells Invest. Ophthalmol. Vis. Sci., April 1, 2005; 46(4): 1525 - 1532. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Lemon, J. Brockhausen, G.-H. Li, W. G. Gibson, and M. R. Bennett Calcium Mobilization and Spontaneous Transient Outward Current Characteristics upon Agonist Activation of P2Y2 Receptors in Smooth Muscle Cells Biophys. J., March 1, 2005; 88(3): 1507 - 1523. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. T. Venkatesha, J. Ahamed, C. Nuesch, A. K. Zaidi, and H. Ali Platelet-activating Factor-induced Chemokine Gene Expression Requires NF-{kappa}B Activation and Ca2+/Calcineurin Signaling Pathways: INHIBITION BY RECEPTOR PHOSPHORYLATION AND {beta}-ARRESTIN RECRUITMENT J. Biol. Chem., October 22, 2004; 279(43): 44606 - 44612. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. J. Zwartz, A. Chigaev, D. C. Dwyer, T. D. Foutz, B. S. Edwards, and L. A. Sklar Real-time Analysis of Very Late Antigen-4 Affinity Modulation by Shear J. Biol. Chem., September 10, 2004; 279(37): 38277 - 38286. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Fam, C. J. Gallagher, L. V. Kalia, and M. W. Salter Differential Frequency Dependence of P2Y1- and P2Y2- Mediated Ca 2+ Signaling in Astrocytes J. Neurosci., June 1, 2003; 23(11): 4437 - 4444. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Wang, Y. Zhou, K. Szabo, C. R. Haft, and J. Trejo Down-Regulation of Protease-activated Receptor-1 Is Regulated by Sorting Nexin 1 Mol. Biol. Cell, June 1, 2002; 13(6): 1965 - 1976. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Vassort Adenosine 5'-Triphosphate: a P2-Purinergic Agonist in the Myocardium Physiol Rev, April 1, 2001; 81(2): 767 - 806. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. C. Zambon, R. J. Hughes, J. G. Meszaros, J. J. Wu, B. Torres, L. L. Brunton, and P. A. Insel P2Y2 receptor of MDCK cells: cloning, expression, and cell-specific signaling Am J Physiol Renal Physiol, December 1, 2000; 279(6): F1045 - F1052. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. J. Gu, W. Y. Zhang, L. J. Bendall, I. P. Chessell, G. N. Buell, and J. S. Wiley Expression of P2X7 purinoceptors on human lymphocytes and monocytes: evidence for nonfunctional P2X7 receptors Am J Physiol Cell Physiol, October 1, 2000; 279(4): C1189 - C1197. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Hou, S. Moller, L. Edvinsson, and D. Erlinge Cytokines Induce Upregulation of Vascular P2Y2 Receptors and Increased Mitogenic Responses to UTP and ATP Arterioscler. Thromb. Vasc. Biol., September 1, 2000; 20(9): 2064 - 2069. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. B. Willars, A. Heding, M. Vrecl, R. Sellar, M. Blomenrohr, S. R. Nahorski, and K. A. Eidne Lack of a C-terminal Tail in the Mammalian Gonadotropin-releasing Hormone Receptor Confers Resistance to Agonist-dependent Phosphorylation and Rapid Desensitization J. Biol. Chem., October 15, 1999; 274(42): 30146 - 30153. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. E. Brinson and T. K. Harden Differential Regulation of the Uridine Nucleotide-activated P2Y4 and P2Y6 Receptors. SER-333 AND SER-334 IN THE CARBOXYL TERMINUS ARE INVOLVED IN AGONIST-DEPENDENT PHOSPHORYLATION DESENSITIZATION AND INTERNALIZATION OF THE P2Y4 RECEPTOR J. Biol. Chem., April 6, 2001; 276(15): 11939 - 11948. [Abstract] [Full Text] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
