A simple filtration assay for measuring hydrogen exchange kinetics of proteins: application to peptide and ligand binding reactions.
A simple filtration assay for measuring hydrogen exchange kinetics of proteins: application to peptide and ligand binding reactions.
复制标题
用于测量蛋白质氢交换动力学的简单过滤测定：应用于肽和配体结合反应。
DOI：
10.1016/0003-2697(77)90523-1
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发表时间：
1977
影响因子：
2.9
通讯作者：
A. Schreier
中科院分区：
文献类型：
--
作者：
A. Schreier
描述了用于测量蛋白质和肽的氢交换动力学的快速过滤测定法。在此测定中，氚标记的蛋白质或肽通过吸附到 pH 3、0°C 的磷酸纤维素纸上而与氚化溶剂分离。该过滤测定产生的数据与通过凝胶过滤和冻干测定获得的数据相同。通过测量与 RNase S 结合的 S 肽（8.8 × 10−11m，pH 6.9，0°C）和与 RNase S 结合的 2'-CMP（4 × 10−7m，pH 6.4，0°C）的小解离常数的交换数据，证明了该测定的实用性。
A rapid filter assay for measurement of the hydrogen exchange kinetics of proteins and peptides is described. In this assay a tritium-labeled protein or peptide is separated from tritiated solvent by adsorption onto phosphocellulose paper at pH 3, 0°C. This filter assay yields data identical to data obtained by gel filtration and lyophilization assays. The utility of the assay is demonstrated by the measurement from exchange data of the small dissociation constants of S-peptide bound to RNase S (8.8 × 10−11m at pH 6.9, 0°C) and of 2′-CMP bound to RNase S (4 × 10−7m at pH 6.4, 0°C).