Modulating effect of adenosine deaminase on function of adenosine A 1 receptors 1
dc.contributor.author | Sun, Wan-Chun | en_US |
dc.contributor.author | Cao, Yan | en_US |
dc.contributor.author | Jin, Lei | en_US |
dc.contributor.author | Wang, Li-Zhen | en_US |
dc.contributor.author | Meng, Fan | en_US |
dc.contributor.author | Zhu, Xing-Zu | en_US |
dc.date.accessioned | 2010-06-01T20:35:08Z | |
dc.date.available | 2010-06-01T20:35:08Z | |
dc.date.issued | 2005-02 | en_US |
dc.identifier.citation | SUN, Wan-chun; CAO, Yan; JIN, Lei; WANG, Li-zhen; MENG, Fan; ZHU, Xing-zu (2005). "Modulating effect of adenosine deaminase on function of adenosine A 1 receptors 1 ." Acta Pharmacologica Sinica 26(2): 160-165. <http://hdl.handle.net/2027.42/73693> | en_US |
dc.identifier.issn | 1671-4083 | en_US |
dc.identifier.issn | 1745-7254 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/73693 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=15663892&dopt=citation | en_US |
dc.description.abstract | Aim : To study the modulating effect of adenosine deaminase (ADA) on yhe adenosine A 1 receptor (A 1 R) in HEK293 cells stably expressing the human A 1 R. Methods : cDNA was amplified by RT-PCR using total RNA from human embryo brain tissue as the template. The PCR products were subcloned into the plasmid pcDNA3 and cloned into the plasmid pcDNA3.1. The cloned A 1 R cDNA was sequenced and stably expressed in HEK293 cells. The modulating effect of adenosine deaminase on A 1 R was studied by using [ 3 H]DPCPX binding assay and an intracellular calcium assay. Results : HEK293 cells stably expressing human A 1 R were obtained. Saturation studies showed that the K D value and B max value of [ 3 H]DPCPX were 1.6±0.2 nmol/L and 1.819±0.215 nmol/g of protein respectively, in the absence of ecto-ADA respectively, and 1.3±0.2 nmol/L and 1.992±0.130 nmol/g of protein in the presence of ecto-ADA respectively, suggesting that the K D value and B max value of [ 3 H]DPCPX were unaffected by ecto-ADA. In the case of [ 3 H]DPCPX competition curves obtained from intact cells or membranes, A 1 R agonist CCPA/[ 3 H]DPCPX competition curve could be fitted well to a one-site model in the absence of ecto-ADA and a two-site model in the presence of ecto-ADA with a K H value of 0.74 (0.11–4.8) nmol/L (intact cells) or 1.8 (0.25–10) nmol/L (membrane) and a K L value of 0.94 (0.62–1.41) Μmol/L (intact cells) or 0.77 (0.29–0.99) Μmol/L (membrane). The K L value is not significantly different from the EC 50 value of 0.84(0.57–1.23) Μmol/L (intact cells) or 0.84 (0.63–1.12) Μmol/L (membrane) obtained in the absence of ecto-ADA. Similar results were obtained from the CPA/[ 3 H]DPCPX competition curve in the absence or presence of ecto-ADA on intact cells or membranes. Intracellular calcium assay demonstrated that the EC 50 value of CPA were 10 (5–29) nmol/L and 94 (38–229) nmol/L in the presence or absence of ecto-ADA, respectively. Conclusion : A 1 R stably expressed in the HEK293 cells display a low affinity for agonists in the absence of ADA and high and low affinities for agonists in the presence of ADA. The presence of ADA may promote the signaling through the adenosine A 1 receptor in HEK293 cells. | en_US |
dc.format.extent | 450258 bytes | |
dc.format.extent | 3109 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.publisher | Blackwell Science Pty | en_US |
dc.rights | 2005 CPS and SIMM | en_US |
dc.subject.other | Adenosine Deaminase | en_US |
dc.subject.other | Adenosine a 1 Receptor | en_US |
dc.subject.other | Radioligand Assay | en_US |
dc.subject.other | Calcium | en_US |
dc.subject.other | Fluorescence | en_US |
dc.subject.other | Diagnosis | en_US |
dc.title | Modulating effect of adenosine deaminase on function of adenosine A 1 receptors 1 | en_US |
dc.type | Article | en_US |
dc.subject.hlbsecondlevel | Pharmacy and Pharmacology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Psychiatry, University of Michigan, The Psychiatry/MHRI Microarray Laboratory, 205 Zina Pitcher Place, Ann Arbor, MI 48109-0720, USA | en_US |
dc.contributor.affiliationother | Department of Pharmacology, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203, China; | en_US |
dc.identifier.pmid | 15663892 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/73693/1/j.1745-7254.2005.00524.x.pdf | |
dc.identifier.doi | 10.1111/j.1745-7254.2005.00524.x | en_US |
dc.identifier.source | Acta Pharmacologica Sinica | en_US |
dc.identifier.citedreference | Wlliams M. Purine receptors in mammalian tissues:pharmacology and functional significance. Annu Rev Pharmacol Toxicol 1987; 27: 315 – 45. | en_US |
dc.identifier.citedreference | Ralevic V, Burnstock G. Receptors for purines and pyrimidines. Pharmacol Review 1998; 50: 413 – 92. | en_US |
dc.identifier.citedreference | Ren H, Stiles G L. Characterization of the human A 1 adenosine receptor gene: evidence for alternative splicing. J Biol Chem 1994; 269: 3104 – 10. | en_US |
dc.identifier.citedreference | Amoah-Apraku B, Xu J, Lu JY, Pelleg A, Bruns RF, Belardinelli L. Selective potentiation by an A 1 adenosine receptor enhancer of the negative dromotropic action of adenosine in the guinea pig heart. J Pharmacol Exp Ther 1993; 266: 611 – 7. | en_US |
dc.identifier.citedreference | Spielman WS, Arend LJ. Adenosine receptors and signaling in the kidney. Hypertension 1991; 17: 117 – 30. | en_US |
dc.identifier.citedreference | el-Hashim AD, Agostino B, Matera MG, Page C. Characterization of a denosine receptors involved in a denosine-induced bronchoconstriction in allergic rabbits. Br J Pharmacol 1996; 119: 1262 – 8. | en_US |
dc.identifier.citedreference | Nyce JW, Metzger WJ. DNA antisense therapy for asthma in an animal model. Nature 1997; 385: 721 – 5. | en_US |
dc.identifier.citedreference | Munshi R, Pang IH, Sternweis PC, Linden J. A 1 adenosine receptors of bovine brain couple to guanine nucleotide-binding proteins Gi1, Gi2 and Go. J Biol Chem 1991; 266: 22285 – 9. | en_US |
dc.identifier.citedreference | Jockers R, Linder ME, Hohenegger M, Nanoff C, Bertin B, Strosberg AD, et al. Species difference in the G protein selectivity of the hu man and bovine A1-adenosine receptor. J Biol Chem 1994; 269: 32077 – 84. | en_US |
dc.identifier.citedreference | Figler RA, Graber SG, Lindorfer MA, Yasuda H, Linden J, Garrison JC. Reconstitution of recombinant bovine A 1 adenosine receptors in Sf9 cell membranes with recombinant G proteins of defined composition. Mol Pharmacol 1996; 50: 1587 – 95. | en_US |
dc.identifier.citedreference | Londos C, Cooper DM, Wolff J. Subclasses of external adenosine receptors. Proc Natl Acad Sci USA 1980; 77: 2551 – 4. | en_US |
dc.identifier.citedreference | Palmer TM, Stiles GL. Adenosine receptors. Neuropharmacology 1995; 34: 683 – 94. | en_US |
dc.identifier.citedreference | Lohse MJ, Lenschow V, Schwabe U. Two affinity states of Ri adenosine receptors in brain membranes. Analysis of guanine nucleotide and temperature effects on radioligand binding. Mol Pharmacol 1984; 26: 1 – 9. | en_US |
dc.identifier.citedreference | Casado V, Canti C, Mallol J, Canela EI, Lluis C, Franco R. Solubilization of A 1 adenosine receptor from pig brain: characterization and evidence of the role of the cell membrane on the coexistence of high and low-affinity states. J Neurosci Res 1990; 26: 461 – 73. | en_US |
dc.identifier.citedreference | Ciruela F, Casado V, Mallol J, Canela EI, Lluis C, Franco R. Immunological identification of A 1 adenosine receptors in brain cortex. J Neurosci Res 1995; 42: 818 – 28. | en_US |
dc.identifier.citedreference | Saura C, Ciruela F, Casado V, Canela EI, Mallol J, Lluis C, et al. Adenosine deaminase interacts with A 1 adenosine receptors in pig brain cortical membranes. J Neurochem 1996; 66: 1675 – 82. | en_US |
dc.identifier.citedreference | Saura CA, Mallol J, Canela EI, Lluis C, Franco R. Adenosine deaminase and A 1 adenosine receptors internalize together following agonist-induced receptor desensitization. J Biol Chem 1998; 273: 17610 – 7. | en_US |
dc.identifier.citedreference | Sorensen K, Brodbeck U. Assessment of coating-efficiency in ELISA plates by direct protein determination. J Immunol Methods 1986; 95: 291 – 3. | en_US |
dc.identifier.citedreference | Sun WC, Jin L, Cao Y, Wang LZ, Meng F, Zhu XZ. Cloning, expression, and functional analysis of human dopamine d1 receptors. Acta Pharmacol Sin 2005; 26: 27 – 32. | en_US |
dc.identifier.citedreference | Kassack MU, Hofgen B, Lehmann J, Eckstein N, Quillan JM, Sadee W. Functional screening of G protein-coupled receptors by measuring intracellular calcium with a fluorescence microplate reader. J Biomol Screen 2002; 7: 233 – 46. | en_US |
dc.identifier.citedreference | Franco R, Casado V, Ciruela F, Saura C, Mallol J, Canela EI, et al. Cell surface adenosine deaminase: much more than an ectoenzyme. Prog Neurobiol 1997; 52: 283 – 94. | en_US |
dc.identifier.citedreference | Ciruela F, Saura C, Canela EI, Mallol J, Lluis C, Franco R. Adenosine deaminase affects ligand-induced signalling by interacting with cell surface adenosine receptors. FEBS Lett 1996; 380: 219 – 23. | en_US |
dc.identifier.citedreference | Escriche M, Burgueno J, Ciruela F, Canela EI, Mallol J, Enrich C, et al. Ligand-induced caveolae-mediated internalization of A 1 adenosine receptors: morphological evidence of endosomal sorting and receptor recycling. Exp Cell Res 2003; 285: 72 – 90. | en_US |
dc.identifier.citedreference | Cohen FR, Lazareno S, Birdsall NJ. The affinity of adenosine for the high-and low-affinity states of the human adenosine A 1 receptor. Eur J Pharmacol 1996; 309: 111 – 4. | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
Files in this item
Remediation of Harmful Language
The University of Michigan Library aims to describe library materials in a way that respects the people and communities who create, use, and are represented in our collections. Report harmful or offensive language in catalog records, finding aids, or elsewhere in our collections anonymously through our metadata feedback form. More information at Remediation of Harmful Language.
Accessibility
If you are unable to use this file in its current format, please select the Contact Us link and we can modify it to make it more accessible to you.