Developement of abiwave stopped flow fluorophotometer

得到以下结果：1、该仪器允许在0.12ml的样品体积下跟踪两种不同的荧光变化，时间尺度为50毫秒到30秒。2、ATP不与E1P结合，加速磷酸酶3的降解，BIPM探针的结合部位为Na~+泵α链的Cys 964。4、硫代荧光探针之间的荧光能量传递发生了异相变化，表现为Na~(++)、K~(++)在主动转运过程中的动态变化。5、Na~(2+)、Na~(2+)和ATP与Na~+泵结合引起构象变化。寡霉素阻断Na~+也引起类似的变化。这些数据有力地表明，镁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^+.