钠通道的分离纯化及其结合活性的研究  

Study on the purification and binding activity of the sodium channel

作　　者：李勇锋[1] 刘敏[1] 

机构地区：[1]防化指挥工程学院生物防护教研室,北京102205

出　　处：《中国药理学通报》2007年第11期1518-1522,共5页Chinese Pharmacological Bulletin

摘　　要：目的对大鼠脑和骨骼肌纤维膜进行钠离子通道分离和纯化,并将其应用于芋螺毒素的研究。方法3种色谱分离纯化钠通道粗组分,配体结合实验研究钠通道活性及其与桶型芋螺毒素的相互作用。结果经过纯化,大鼠脑钠通道纯化倍数达370倍,平均特异活性位点数达1.3nmol.g-1。大鼠骨骼肌钠通道被纯化2436倍,平均特异活性位点数达268nmol.g-1;用纯化的钠通道与桶型芋螺毒素进行竞争结合,发现Ⅴ8成分与钠通道作用最强。结论纯化出了较高纯度的大鼠脑钠通道和大鼠骨骼肌钠通道;桶型芋螺毒素中的Ⅴ8成分与钠通道作用强烈,与传统型μ-芋螺毒素非常相似。Aim To apply sodium channels isolated and purified from rat brain and rat skelatal muscle sarcolemma to the study conotoxin.Methods Crude isolations of sodium channels were purified by 3 kinds of chromatography.The interaction between the sodium channels and conus betulinus was studied by the ligand binding experiment.Result The sequential chromatography resulted in 370-fold purification from rat brain and 2436-fold purification from rat skeletal muscle.Average specific binding active sites of purified sodium channels increased to 321 nmol·g^-1 for rat brain and 268 nmol·g^-1 for rat skeletal muscle respectively;Studying the interaction between venom from conus betulinus and the purified sodium channels indicated the fraction Ⅴ8 showed the highest affinity for the binding sites of the purified sodium channels.Conclusion Rat brain sodium channel and rat skeletal muscle sodium channel were purified to high purity;the fraction Ⅴ8 has the similar effect to that of the traditional μ-conotoxin.

关 键 词：钠通道 桶型芋螺毒素 色谱纯化 结合实验 

分 类 号：R-332[医药卫生] R322.81