人神经母细胞瘤株SH-SY5Y中calcineurin比活性测定的最佳条件  

OPTIMIZING CONDITIONS FOR CALCINEURIN ACTIVITY IN SH-SY5Y CELL EXTRACTS

作　　者：涂玲辉[1] 马杰 佟丽[1] 魏群[1] 骆静[1] 

机构地区：[1]北京师范大学生命科学学院,北京市基因工程药物及生物技术重点实验室,北京100875

出　　处：《北京师范大学学报（自然科学版）》2009年第1期66-69,共4页Journal of Beijing Normal University(Natural Science)

基　　金：国家自然科学基金资助项目(G30772558,G30470393,J0630642)

摘　　要：研究了在人神经母细胞瘤株SH-SY5Y中calcineirin比活性测定的最佳条件.以RⅡ为底物,在测活缓冲液中,Ca2+和岗田酸(okadaic acid,OA)的加入是必须的,这与在脑组织中的测定是不一样的,同时进一步确定了需要加入的OA是100 nmol.L-1.在细胞破碎液中,calcineurin比活性的测定值是以缓冲液Ⅰ(含Ca2+与OA)中测得CN比活性数值减去缓冲液Ⅱ(含EGTA与OA)中所测比活性数值之差来表示.Effects of Ca^2＋ and OA on calcineurin （CN） activity in SH-SY5Y cell extracts were investigated with RⅡ phosphopeptide as substrate. Data showed that adding Ca^2＋ to assay buffer Ⅰ was necessary to activate CN in SH-SY5Y cell extracts, which was unlike extracts from brain. Further, it was necessary to add OA to assay buffers to inhibit PP1 and PP2A in SH-SY5Y cell extracts, with an optimal OA concentration of 100 nmol.L^-1, which was also unlike the brain extracts. In conclusion, calcineurin phosphatase activity in SH-SY5Y cell extracts should be calculated as the difference in protein phosphatase activity in buffer Ⅰ （cantains Ca^2＋ and OA） and buffer Ⅱ （cantains EGTAand OA）＂.

关 键 词：钙调神经磷酸酶 人神经母细胞瘤株 RⅡ多肽 

分 类 号：Q78[生物学—分子生物学] S858.31[农业科学—临床兽医学]