Recommendations for the Propagation of Uncertainty in Hydrogen Exchange-Mass Spectrometric Measurements
Recommendations for the Propagation of Uncertainty in Hydrogen Exchange-Mass Spectrometric Measurements
复制标题
关于氢交换质谱测量中不确定性传播的建议
DOI：
10.1021/jasms.0c00475
复制
发表时间：
2021
影响因子：
3.2
通讯作者：
Weis, David D.
中科院分区：
文献类型：
--
作者：
Weis, David D.
氢交换-质谱仪(HX-MS)被广泛用于表征蛋白质的高阶结构，并定位伴随着配体结合和蛋白质-蛋白质相互作用的蛋白质结构和动力学的变化。两个态之间氢交换量的数量差异(即差值HX)被认为是高阶结构或动力学存在显著差异的证据。定量测量的范围从一个HX标记时间的简单质量差异到整个HX时间过程中的平均差异，并修正了氘的回收。这项工作将不确定度传播原理应用于差分HX测量，以便于识别显著差异。此外，实验不确定度的合并估计比基于单个标准差的不确定度估计的假阳性率更低。
Hydrogen exchange-mass spectrometry (HX-MS) is used widely to characterize higher-order protein structure and to locate changes in protein structure and dynamics that accompany, for example, ligand binding and protein–protein interactions. Quantitative differences in the amount of hydrogen exchange between two states (i.e., differential HX) are taken as evidence of significant differences in higher-order structure or dynamics. The quantitative measures range from simple mass differences at one HX labeling time to differences averaged across an HX time course with correction for deuterium recovery. This work applies the principles of uncertainty propagation to differential HX measurements to facilitate the identification of significant differences. Furthermore, it is shown that pooled estimates of experimental uncertainty result in a lower false positive rate than estimates of uncertainty based on individual standard deviations.