Developement of abiwave stopped flow fluorophotometer

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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^+.