Studies of Chemically Reacting Systems on Sephadex. II. Molecular Weights of Monomers in Rapid Association Equilibrium
Studies of Chemically Reacting Systems on Sephadex. II. Molecular Weights of Monomers in Rapid Association Equilibrium
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Sephadex 化学反应系统的研究。
DOI：
10.1021/j100784a022
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发表时间：
1964
期刊：
影响因子：
--
通讯作者：
H. Scheraga
中科院分区：
文献类型：
--
作者：
D. Winzor;H. Scheraga
在对经历快速、可逆缔合的蛋白质的研究中，由于需要将实验数据外推至零浓度，因此通过物理化学方法估计单体分子量变得困难。因此，这种测定的准确性在很大程度上取决于可通过特定方法研究的浓度范围的极限。研究浓度低至 0.05 mg./ml 的蛋白质溶液已成为可能。通过采用凝胶过滤的程序，其中首先确定分子量和洗脱速率之间的关系。已使用醋酸盐缓冲液（离子强度 0.20，pH 3.86）中的乳糜蛋白酶研究来检查该程序的有效性，然后将其应用于磷酸盐缓冲液（离子强度 0.16，pH 7.0）中牛凝血酶单体分子量的估计；在这些条件下获得的值约为 40,000。
In studies of proteins undergoing rapid, reversible association, estimation of the monomeric molecular weights by physicochemical methods is made difficult bythe necessity of extrapolating the experimental data to zero concentration. Thus, the accuracy of such de-terminations depends to a large extent upon the knver limit of the concentration range which can be investigated by the particular method. It has been possible to study protein solutions with concentrations as low as 0.05 mg./ml. by a procedure employing gel filtration, in which the relationship between molecularweight and rate of elution is first determined. A study of-chy mo trypsin in acetate-chloride buffer, ionic strength 0.20, pH 3.86, has been used to check the validity of the procedure, which has then been applied to the estimation of the monomeric molecular weight of bovine thrombin in phosphatechloride buffer, ionic strength 0.16, pH 7.0; a value of approximately 40,000 is obtained under these conditions.