【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.更多还原

【Abstract】 Effects of Ca2+ and OA on calcineurin(CN) activity in SH-SY5Y cell extracts were investigated with RⅡ phosphopeptide as substrate.Data showed that adding Ca2+ 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 Ca2+ and OA) and buffer Ⅱ(cantains EGTAand OA)".更多还原