MEASUREMENT AND CALIBRATION OF PEPTIDE GROUP HYDROGEN-DEUTERIUM EXCHANGE BY ULTRAVIOLET SPECTROPHOTOMETRY
MEASUREMENT AND CALIBRATION OF PEPTIDE GROUP HYDROGEN-DEUTERIUM EXCHANGE BY ULTRAVIOLET SPECTROPHOTOMETRY
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DOI：
10.1016/0003-2697(79)90693-6
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发表时间：
1979-01-01
影响因子：
2.9
通讯作者：
ENGLANDER, SW
中科院分区：
文献类型：
--
作者：
ENGLANDER, JJ;CALHOUN, DB;ENGLANDER, SW
本文介绍了一种用紫外分光光度法测定含肽键分子氢氘交换行为的简便方法。交换反应通过将样品从H2O稀释到D2 O或相反来启动，并且随后可以在肽吸光度区域中容易观察到光密度变化。与红外和磁共振方法不同，该方法只需要少量的材料，并且与氚-葡聚糖凝胶方法不同，该方法不限于研究大分子。提供了作为pD（氘离子浓度倒数的对数）和温度函数的交换率以及吸光度/mol肽基团变化的校准。利用该信息，可以预测暴露于溶剂的任何肽基团的预期交换曲线。然后与测量数据进行比较可以识别肽基团H-键合，并且还可以给出H-键合结构的稳定性的量度。
An exceedingly simple and convenient method is described for measuring the H-deuterium exchange behavior of peptide bond-containing molecules by UV spectrophotometry. The exchange reaction is initiated by diluting a sample from H2O into D2O, or the reverse, and can be followed by an easily observable optical density change in the region of peptide absorbance. The method, unlike IR and magnetic resonance approaches, requires only small amounts of material and, unlike the tritium-Sephadex method, is not restricted to the study of large molecules. Calibrations are provided for exchange rate as a function of pD (log of the reciprocal of the deuterium ion concentration) and temperature and for the change in absorbance/mol peptide group. With this information, the exchange curve to be expected for any peptide group exposed to solvent can be predicted. Comparison with the measured data can then identify peptide-group H-bonding and can also give a measure of the stability of the H-bonded structure.