Proteome dynamics from heavy water metabolic labeling and peptide tandem mass spectrometry
Proteome dynamics from heavy water metabolic labeling and peptide tandem mass spectrometry
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DOI：
10.1016/j.ijms.2019.116194
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发表时间：
2019-11-01
影响因子：
1.8
通讯作者：
Sadygov, Rovshan G.
中科院分区：
文献类型：
--
作者：
Borzou, Ahmad;Sadygov, Vugar R.;Sadygov, Rovshan G.
蛋白质稳态（Proteostasis）是蛋白质合成和降解之间动态平衡的结果。它对健康的细胞/器官功能很重要，通常与神经退行性疾病和非酒精性脂肪肝等疾病有关。重水代谢标记结合液相色谱-质谱联用技术是高通量研究体内蛋白质代谢稳定性的有效方法。传统上，完整的肽信号被用来估计稳定的同位素掺入的时间过程实验。利用标记掺入的时间过程提取蛋白质衰减速率常数（DRC）。完整的肽信号，从色谱时间和质荷比（m/z）域的积分计算，通常，提供了一个准确的估计标签掺入。然而，样品复杂性（共洗脱）、有限的动态范围和低信噪比（S/N）可能不利地干扰肽信号。这些伪影通过使色谱时间和m/z域中的峰形失真而使DRC估计复杂化。另一方面，碎片离子不太容易出现这些伪影，并且可能非常适合于辅助DRC估计。在这里，我们表明，标签纳入编码到同位素分布的碎片离子反映同位素富集的代谢标记与重水。我们探讨标签纳入统计设计实用的方法DRC估计。由爱思唯尔公司出版
Protein homeostasis (proteostasis) is a result of a dynamic equilibrium between protein synthesis and degradation. It is important for healthy cell/organ functioning and is often associated with diseases such as neurodegenerative diseases and non-Alcoholic Fatty Liver disease. Heavy water metabolic labeling, combined with liquid-chromatography and mass spectrometry (LC-MS), is a powerful approach to study proteostasis in vivo in high throughput. Traditionally, intact peptide signals are used to estimate stable isotope incorporation in time-course experiments. The time-course of label incorporation is used to extract protein decay rate constant (DRC). Intact peptide signals, computed from integration in chromatographic time and mass-to-charge ratio (m/z) domains, usually, provide an accurate estimate of label incorporation. However, sample complexity (co-elution), limited dynamic range, and low signal-to-noise ratio (S/N) may adversely interfere with the peptide signals. These artifacts complicate the DRC estimations by distorting peak shape in chromatographic time and m/z domains. Fragment ions, on the other hand, are less prone to these artifacts and are potentially well suited in aiding DRC estimations. Here, we show that the label incorporation encoded into the isotope distributions of fragment ions reflect the isotope enrichment during the metabolic labeling with heavy water. We explore the label incorporation statistics for devising practical approaches for DRC estimations. Published by Elsevier B.V.