Modulating effect of adenosine deaminase on function of adenosine A1 receptors
文献类型：期刊论文
| 作者 | Sun Wanchun2; Cao Yan2; Jin Lei2; Wang Lizhen2; Meng Fan1; Zhu Xingzu1 |
| 刊名 | 中国药理学报：英文版
 |
| 出版日期 | 2005 |
| 卷号 | 26.0期号:002页码:160 |
| 关键词 | 调节作用 腺甙脱氨酶 腺苷A1受体 放射配体测定 荧光性 |
| ISSN号 | 1671-4083 |
| 英文摘要 | Aim: To study the modulating effect of adenosine deaminase (ADA) on yhe adenosine AI receptor (AIR) in HEK293 cells stably expressing the human A1R. 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 AIR cDNA was sequenced and stably expressed in HEK293 cells. The modulating effect of adenosine deaminase on AIR was studied by using 3HDPCPX binding assay and an intracellular calcium assay. Results: HEK293 cells stably expressing human AIR were obtained. Saturation studies showed that the KD value and Bmax value of ^3H 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 KD value and Bmax value of ^3H DPCPX were unaffected by ecto-ADA. In the case of ^3H DPCPX competition curves obtained from intact cells or membranes, A1R agonist CCPA/^3HDPCPX 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 ectoADA with a KH value of 0.74 (0.11-4.8) nmol/L (intact cells) or 1.8 (0.25-10) nmol/L(membrane) and a KL value of 0.94 (0.62-1.41) umol/L (intact cells) or 0.77 (0.29-0.99) umol/L (membrane). The KL value is not significantly different from the IC50 value of 0.84(0.57-1.23) umol/L (intact cells) or 0.84 (0.63-1.12) umol/L (membrane) obtained in the absence of ecto-ADA. Similar results were obtained from the CPA/^3HDPCPX 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: AIR 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 AI receptor in HEK293 cells. |
| 语种 | 英语 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/293938]  |
| 专题 | 中国科学院上海药物研究所 |
| 作者单位 | 1.密歇根大学 2.中国科学院上海药物研究所 |
| 推荐引用方式 GB/T 7714 | Sun Wanchun,Cao Yan,Jin Lei,et al. Modulating effect of adenosine deaminase on function of adenosine A1 receptors[J]. 中国药理学报：英文版,2005,26.0(002):160. |
| APA | Sun Wanchun,Cao Yan,Jin Lei,Wang Lizhen,Meng Fan,&Zhu Xingzu.(2005).Modulating effect of adenosine deaminase on function of adenosine A1 receptors.中国药理学报：英文版,26.0(002),160. |
| MLA | Sun Wanchun,et al."Modulating effect of adenosine deaminase on function of adenosine A1 receptors".中国药理学报：英文版 26.0.002(2005):160. |
入库方式： OAI收割
来源：上海药物研究所
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