Modulating effect of adenosine dearninase on function of adenosine A(1) receptors
文献类型：期刊论文
| 作者 | Sun, WC; Cao, Y; Jin, L; Wang, LZ; Meng, F; Zhu, XZ |
| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2005-02 |
| 卷号 | 26期号:2页码:160-165 |
| 关键词 | adenosine deaminase adenosine A(1) receptor radioligand assay calcium fluorescence diagnosis |
| ISSN号 | 1671-4083 |
| DOI | 10.1111/j.1745-7254.2005.00024.x |
| 文献子类 | Article |
| 英文摘要 | 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,R cDNA was sequenced and stably expressed in HEK293 cells. The modulating effect of adenosine deaminase on A(1)R was studied by using [H-3]DPCPX binding assay and an intracellular calcium assay. Results: HEK293 cells stably expressing human A,R were obtained. Saturation studies showed that the K-D value and B-max value of [H-3]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 [H-3]DPCPX were unaffected by ecto-ADA. In the case of [3 H]DPCPX competition curves obtained from intact cells or membranes, A,R agonist CCPA/[H-3]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, value of 0.74 (0.11-4.8) nmol/L (intact cells) or 1.8 (0.25-10) nmol/L (membrane) and a K-H value of 0.94 (0.62-1.4 1) mumol/L (intact cells) or 0.77 (0.29-0.99) mumol/L (membrane). The K-L value is not significantly different from the IC50 value of 0.84(0.57-1.23) mumol/L (intactcells) or 0.84 (0.63-1.12) mumol/L (membrane) obtained in the absence of ecto-ADA. Similar results were obtained from the CPA/[H-3]DPCPX competition curve in the absence or presence of ecto-ADA on intact cells or membranes. Intracellular calcium assay demonstrated that the EC50 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,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, receptor in HEK293 cells. |
| WOS关键词 | LOW-AFFINITY STATES ; A(1)-ADENOSINE RECEPTOR ; GUANINE-NUCLEOTIDE ; PIG BRAIN ; DEAMINASE ; MEMBRANES ; PROTEINS ; BINDING |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:1918235 |
| WOS记录号 | WOS:000226942000005 |
| 出版者 | ACTA PHARMACOLOGICA SINICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/273912]  |
| 专题 | 药理学第二研究室 |
| 通讯作者 | Zhu, XZ |
| 作者单位 | 1.Univ Michigan, Dept Psychiat, Psychiat MHRI Microarray Lab, Ann Arbor, MI 48109 USA 2.Chinese Acad Sci, Shanghai Inst Biol Sci, Dept Pharmacol, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Sun, WC,Cao, Y,Jin, L,et al. Modulating effect of adenosine dearninase on function of adenosine A(1) receptors[J]. ACTA PHARMACOLOGICA SINICA,2005,26(2):160-165. |
| APA | Sun, WC,Cao, Y,Jin, L,Wang, LZ,Meng, F,&Zhu, XZ.(2005).Modulating effect of adenosine dearninase on function of adenosine A(1) receptors.ACTA PHARMACOLOGICA SINICA,26(2),160-165. |
| MLA | Sun, WC,et al."Modulating effect of adenosine dearninase on function of adenosine A(1) receptors".ACTA PHARMACOLOGICA SINICA 26.2(2005):160-165. |
入库方式： OAI收割
来源：上海药物研究所
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