Quantitating the statistical distribution of deuterium incorporation to extend the utility of H/D exchange MS data
Quantitating the statistical distribution of deuterium incorporation to extend the utility of H/D exchange MS data
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DOI：
10.1021/ac050988l
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发表时间：
2006-01-01
影响因子：
7.4
通讯作者：
Schriemer, DC
中科院分区：
文献类型：
--
作者：
Chik, JK;Graaf, JLV;Schriemer, DC
测量掺入酶促衍生的肽片段中的氘的统计分布为氢/氘交换质谱数据提供了有价值的维度。在本文中，我们将讨论我们的线性最小二乘法确定此分布的改进，通过添加“零”的氘同位素信封的末端，部分补偿由于有限的仪器信噪比的数据截断。分布的值在一个简单的实验示例中得到证明，其中用于标记测试肽的平均氘代和D2 O百分比之间的线性隐藏了位点标记概率和位点总数之间更复杂的关系。这种方法提供了解决单个肽经历不同的、独立的生化状态的情况的机会，每个肽都具有独特的平均氘代;当蛋白质被修饰到亚化学计量水平时，可能会发生这种情况。从实验确定的分布的不均匀氘化肽，它是可能的，以提取平均氘化的混合物的每一个组成部分。
Measuring the statistical distribution of deuterium incorporated into enzymatically derived peptide fragments provides a valuable dimension to hydrogen/deuterium exchange mass spectrometry data. In this paper, we will discuss our improvement to the linear least-squares method for determining this distribution, through the addition of "zeroes" to the end of the deuterated isotopic envelope, to partially compensate for data truncation due to finite instrumental signal-to-noise ratios. The value of the distribution is demonstrated in a simple experimental example, where the linearity between average deuteration and percent D2O used to label test peptides hides a more complex relationship between the site-labeling probability and the total number of sites. This method offers the opportunity to resolve cases where a single peptide experiences distinct, independent biochemical states with each beating a unique average deuteration; this can occur when a protein is modified to substoichiometric levels. From the experimentally determined distribution of a heterogeneously deuterated peptide, it was possible to extract the average deuteration of each component of the mixture.