AGGREGATION OF BOVINE SERUM-ALBUMIN UPON CLEAVAGE OF ITS DISULFIDE BONDS, STUDIED BY THE TIME-RESOLVED SMALL-ANGLE X-RAY-SCATTERING TECHNIQUE WITH SYNCHROTRON RADIATION
AGGREGATION OF BOVINE SERUM-ALBUMIN UPON CLEAVAGE OF ITS DISULFIDE BONDS, STUDIED BY THE TIME-RESOLVED SMALL-ANGLE X-RAY-SCATTERING TECHNIQUE WITH SYNCHROTRON RADIATION
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DOI：
10.1016/0301-4622(85)80069-7
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发表时间：
1985-11-01
影响因子：
3.8
通讯作者：
MUROGA, Y
中科院分区：
文献类型：
--
作者：
UEKI, T;HIRAGI, Y;MUROGA, Y
介绍了一种与同步辐射小角度x射线散射设备配套使用的停止流型快速混合系统。牛血清白蛋白的聚集过程以50 s的时间间隔进行了追踪，该过程是由二硫苏糖醇还原其二硫键的断裂开始的。结果表明，双硫苏糖醇的摩尔量超过牛血清白蛋白中二硫键摩尔数的218倍，足以在混合后立即引发反应，并在约15分钟内达到平衡。另一方面，这个量的一半不足以引起反应，因此反应延迟约150秒。这种单镜头时间分辨实验表明，在重复的多镜头运行中，时间间隔为100 ms的实验是可能的。
A rapid mixing system of the stopped-flow type, used with small-angle X-ray scattering equipment using synchrotron radiation, is described. The process of aggregation of bovine serum albumin was traced with a time interval of 50 s, initiated upon cleavage of its disulfide bonds by reduction with dithiothreitol. The results indicate that a 218-fold molar excess of dithiothreitol over the number of moles of disulfide bonds in bovine serum albumin is sufficient to initiate the reaction immediately after mixing, which reaches equilibrium in about 15 min. On the other hand, half this amount is not sufficient to initiate the reaction, so that the reaction is delayed by about 150 s. Such a single-shot time-resolved experiment showed that experiments with a time interval of 100 ms are possible with repeated multi-shot runs.