Developement of abiwave stopped flow fluorophotometer

获得以下结果1，仪器允许用0.12ml样品体积跟踪两种不同的荧光变化，时间尺度为50毫秒至30秒。2、ATP不与E1P结合，从而加速磷酸酶的分解。3、BIPM探针的结合位点为Na^+泵α链的Cys 964。4、巯基荧光探针间荧光能量传递的变化是异相的，表现为Na^+、K^+主动转运过程中构象的动态变化。5、Mg^<2+>、Na^+和ATP与Na^+泵的结合引起构象变化。寡霉素对Na^+的封闭作用也引起了类似的变化。这些数据有力地表明，Mg ATP的作用是封闭Na^+。

Following results were obtained 1,The instrument permits to follow two different fluorescence changes with time scale of 50 msec to up to 30 sec with 0.12ml of the sample volume. 2,ATP did no bind to E1P in such a way as to accelerate the breakdown of the phosphoenzyme 3,The site of attachment of BIPM probe was shown to be Cys 964 of the alpha chain of Na^+ pump. 4,Changes in fluorescence energy transfer between sulfhydyl fluorescence probes occured out of phase, which showed dynamic confotmational changes during active transport of Na^+ and K^+. 5,Binding of Mg^<2+>, Na^+ and ATP to Na^+ pump induced the conformational change. Na^+ occlusion by oligomycin also induced the similar change. These data strongly suggest that the role of Mg ATP is to occlude Na^+.