Improved Sequence Resolution by Global Analysis of Overlapped Peptides in Hydrogen/Deuterium Exchange Mass Spectrometry
Improved Sequence Resolution by Global Analysis of Overlapped Peptides in Hydrogen/Deuterium Exchange Mass Spectrometry
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DOI：
10.1007/s13361-012-0373-3
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发表时间：
2012-07-01
影响因子：
3.2
通讯作者：
Marshall, Alan G.
中科院分区：
文献类型：
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作者：
Fajer, Piotr G.;Bou-Assaf, George M.;Marshall, Alan G.
通过质谱监测液相氢/氘交换过程中产生的大量数据的管理刺激了软件分析的发展。实验的蛋白水解步骤产生多个肽片段，其中大部分重叠。先前的自动数据简化算法提取单个肽的氘化水平，但不利用由片段重叠产生的额外信息。在这里，我们描述了一种算法，确定离散的速率常数值的每个重叠片段中的酰胺氢。通过同时考虑所有重叠的肽段，可以显著提高序列分辨率，有时甚至提高到单个氨基酸水平。我们已经验证了该方法与模拟氘摄取数据的聚-丙氨酸九肽的7个重叠片段，然后将其应用于提取速率常数值的前29个N-末端氨基酸的C22 A FK 506结合蛋白。
Management of the enormous amount of data produced during solution-phase hydrogen/deuterium exchange monitored by mass spectrometry has stimulated software analysis development. The proteolysis step of the experiment generates multiple peptide fragments, most of which overlap. Prior automated data reduction algorithms extract the deuteration level for individual peptides, but do not exploit the additional information arising from fragment overlap. Here, we describe an algorithm that determines discrete rate constant values to each of the amide hydrogens in overlapped fragments. By considering all of the overlapped peptide segments simultaneously, sequence resolution can be improved significantly, sometimes to the individual amino acid level. We have validated the method with simulated deuterium uptake data for seven overlapped fragments of a poly-Ala nonapeptide, and then applied it to extract rate constant values for the first 29 N-terminal amino acids of C22A FK506-binding protein.