SEPARATION OF NEUTRAL PROTEINS ON ION-EXCHANGE RESINS
SEPARATION OF NEUTRAL PROTEINS ON ION-EXCHANGE RESINS
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DOI：
10.1042/bj0590543
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发表时间：
1955-01-01
影响因子：
4.1
通讯作者：
PARTRIDGE, SM
中科院分区：
文献类型：
--
作者：
BOARDMAN, NK;PARTRIDGE, SM
研究了缓冲液的pH值和阳离子浓度对IRC-50离子交换树脂吸附细胞色素c的影响。蛋白质在树脂上的行为与低分子量的两性电解质如碱性氨基酸的行为明显不同。当pH值福尔斯降至5以下时，由于树脂羧基的离子化受到抑制，赖氨酸被解吸，但在酸性条件下，蛋白质的吸收大大增加。有人建议，这种吸收是由于大量增加的二次短程力。在IRC-50柱上进行洗脱层析，分离出一些密切相关的中性蛋白。从羊母体CO血红蛋白和牛CO血红蛋白中分离出羊胎儿CO血红蛋白，从牛高铁血红蛋白中分离出牛CO血红蛋白。血红蛋白的分离强烈依赖于洗脱缓冲液的pH和钠离子浓度。为了获得良好的产量，必须在接近0[度]的温度下工作，并使用新鲜制备的蛋白质。对从柱中洗脱的牛CO血红蛋白进行的试验表明，蛋白质通过柱时未发生变化。来自柱的材料可以容易地结晶。
The effect of pH and cation concentration of buffer on the adsorption of cytochrome c on the ion-exchange resin, IRC-50, was studied in some detail. The behavior of the protein on the resin is markedly different from that of ampholytes of low molecular weight such as the basic amino acids. Lysine is desorbed as the pH falls below 5 owing to suppression of the ionization of the resin carboxyl groups, but the absorption of the protein is greatly increased under acidic conditions. It is suggested that this absorption is due to a large increase in secondary short-range forces. Some closely related neutral proteins were separated by elution chromatography on columns of IRC-50. Sheep fetal CO hemoglobin was separated from sheep maternal CO hemoglobin and from bovine CO hemoglobin and bovine CO hemoglobin from bovine methemoglobin. The separation of the hemoglobins is sharply dependent on the pH and sodium ion concentration of the eluting buffer. To obtain good yields, it is necessary to work at a temperature near 0[degree] and to use freshly prepared proteins. Tests carried out on bovine CO hemoglobin eluted from the column indicated that the protein was unaltered by passage through the column. The material from the column could be readily crystallized.