摘要
目的 探讨骨骼肌型ryanodine受体 (RyR1)放射配基分析法的建立及适宜条件。方法蔗糖梯度离心提取骨骼肌肌浆网膜蛋白质组分 ,用3H ryanodine进行放射配基结合实验 ,观察RyR1与3H ryanodine结合的适宜条件。结果 ① 37℃恒温水浴下 ,0~ 6 0min内 ,3H ryanodine与骨骼肌肌浆网膜蛋白质的结合量迅速增加 ,90min时基本达平衡 ,90~ 12 0min内无明显变化 ;②3H ryanodine在1~ 30nmol·L- 1 ,匀浆蛋白质质量浓度为 0 .4~ 1.0g·L- 1 范围内 ,3H ryanodine与RyR1结合的效果最佳。结论 用3H ryanodine在适宜的条件下可建立RyR1放射配基分析法。
Objective To establish an appropriate condition for measuring the skeletal ryanodine receptor 1 (RyR1) in rabbit skeletal sarcoplasmic reticulum (SR). Methods A fraction of skeletal SR membrane was extracted via sucrose gradient centrifugation. 3H ryanodine was used as a radioligand to assay its binding to the skeletal SR membrane fraction of adult rabbits in various protein concentrations and incubation time. Results ①The ryanodine binding kept increasing at 37 ℃ incubation from 0 to 60 min, and reached the equilibrium at 90 min, then remained with it at 90~120 min. ②The optimum binding was obtained when the 3H ryanodine concentration was 1~30 nmol·L -1 and the membrane fraction concentration ranging from 0.4 to 1.0 g·L -1 . Conclusion 3H ryanodine can be used as a radioligand for analysing the RyR1 under appropriate condition.
出处
《中华核医学杂志》
CAS
CSCD
北大核心
2003年第3期171-173,共3页
Chinese Journal of Nuclear Medicine
基金
国家自然科学基金资助项目 (30 1 70 81 1
39870 70 2 )
江苏省教育厅自然科学基金资助项目 (99KJB32 0 0 2 )