摘要
通过在稳定表达趋化因子受体的细胞系CHO/CXCR1、CHO/CXCR2、CHO/CXCR3、CHO/CXCR4和CHO/CCR5上进行[35S]GTPγS结合实验,研究了Saponin对不同趋化因子受体活化的影响。实验结果表明:(1)对于CHO/CXCR1和CHO/CXCR4,在反应体系中添加10μg/mlSaponin能提高受体G蛋白与[35S]GTPγS的特异性结合,使刺激比率(CPMAgonist/CPMBasal)分别从125%和184%扩大到481%和415%;(2)对于CHO/CCR5,10μg/mlSaponin对受体G蛋白与[35S]GTPγS的特异性结合无影响,刺激比率大小未变;(3)对于CHO/CXCR2和CHO/CXCR3,10μg/mlSaponin降低了受体G蛋白与[35S]GTPγS的特异性结合,使刺激比率分别从171%和168%减小到130%和114%。实验结果表明Saponin是与受体发生作用,对于不同的趋化因子受体,Saponin对受体的活化的影响不同。
The effect of Saponin upon the activation of various chemokine receptors was studied by [^35S]GTPγ/S binding assay on cell lines of CHO/CXCR1, CHO/CXCR2, CHO/CXCR3, CHO/CXCR4 and CHO/CCR5 which stably expressed chemokine receptor CXCR1, CXCR2, CXCR3, CXCR4, and CCRS, respectively. The results indicated that ： （ 1 ） On CHO/CXCR1 and CHO/CXCR4, the addition of 10μg/ml of Saponin to the assay system could facilitate the activation of the receptor and the specific binding of [^35S]GTPγ/S to receptor G-protein complexes, enlarge the stimulation ratio （CPMAgonist/CPMBasal） of [^35S]GTPγ/S binding assay from 125% and 184% to 481% and 415%, respectively; （2）On CHO/CCRS, 10μg/ml of Saponin had no effect on the activation of the receptor, the specific binding of [^35S]GTPγ/S to receptor G-protein and the stimulation ratio; （3）On CHO/CXCR2 and CHO/ CXCR3, 10μg/ml of Saponin inhibited the activation of the receptor and reduced the stimulation ratio from 171% and 168% to 130% and 114%, respectively. It was revealed that Saponin had something to do with receptors but not Gprotein. And it was concluded that the effect of Saponin upon the activation of various chemokine receptors were different.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2006年第10期41-45,共5页
China Biotechnology
