Avoiding H/D Scrambling with Minimal Ion Transmission Loss for HDX-MS/MS-ETD Analysis on a High-Resolution Q-TOF Mass Spectrometer.
Avoiding H/D Scrambling with Minimal Ion Transmission Loss for HDX-MS/MS-ETD Analysis on a High-Resolution Q-TOF Mass Spectrometer.
复制标题
在高分辨率 Q-TOF 质谱仪上进行 HDX-MS/MS-ETD 分析时，以最小的离子传输损失避免 H/D 扰乱。
DOI：
10.1021/acs.analchem.9b05208
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发表时间：
2020
影响因子：
7.4
通讯作者：
T. Jørgensen
中科院分区：
文献类型：
--
作者：
Daniel T Weltz Wollenberg;S. Pengelley;J. Mouritsen;D. Suckau;C. I. Jørgensen;T. Jørgensen
通过质谱 (HDX-MS) 监测氢/氘交换，通过测量在 D2O 中孵育的蛋白质中的时间分辨氘掺入，可以研究蛋白质动力学。在气相中使用基于电子的碎裂，可以以单残留分辨率测量氘的吸收。然而，这种方法的先决条件是溶液相标记在前体碎片之前保存在气相中。因此，必须减少甚至避免分子内氢/氘迁移，这会导致氘标记沿着肽随机化（氢扰乱）。在这里，我们描述了一种优化策略，可将扰乱减少到可忽略的水平，同时最大限度地减少对配备 ETD 和电喷雾电离接口（由玻璃传输毛细管和双离子漏斗组成）的高分辨率 Q-TOF 灵敏度的影响。在我们的策略中，我们将优化范围缩小到两个加速潜力，并通过考虑它们与加扰和传输效率相关的相互依赖性，用一个简单的规则定义了它们的优化。利用这一规则，我们能够使用对扰乱高度敏感的四重带电 P1 肽扰乱探针将扰乱率从 75% 平均减少到 5% 以下，从而导致 33% 的传输损失。为了证明这种方法的适用性，我们探测了细胞色素 c 中某些区域的动态。
Hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) enables the study of protein dynamics by measuring the time-resolved deuterium incorporation into a protein incubated in D2O. Using electron-based fragmentation in the gas phase it is possible to measure deuterium uptake at single-residue resolution. However, a prerequisite for this approach is that the solution-phase labeling is conserved in the gas phase prior to precursor fragmentation. It is therefore essential to reduce or even avoid intramolecular hydrogen/deuterium migration, which causes randomization of the deuterium labels along the peptide (hydrogen scrambling). Here, we describe an optimization strategy for reducing scrambling to a negligible level while minimizing the impact on sensitivity on a high-resolution Q-TOF equipped with ETD and an electrospray ionization interface consisting of a glass transfer capillary followed by a dual ion funnel. In our strategy we narrowed down the optimization to two accelerating potentials, and we defined the optimization of these in a simple rule by accounting for their interdependency in relation to scrambling and transmission efficiency. Using this rule, we were able to reduce scrambling from 75% to below 5% on average using the highly scrambling-sensitive quadruply charged P1 peptide scrambling probe resulting in a minor 33% transmission loss. To demonstrate the applicability of this approach, we probe the dynamics of certain regions in cytochrome c.
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DOI：
10.1016/j.str.2018.08.006
发表时间：
2018-12-04
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Karch KR;Coradin M;Zandarashvili L;Kan ZY;Gerace M;Englander SW;Black BE;Garcia BA
通讯作者：
Garcia BA
DOI：
10.1021/acs.jpcb.6b09358
发表时间：
2017-04-20
期刊：
The journal of physical chemistry. B
影响因子：
--
作者：
Saltzberg DJ;Broughton HB;Pellarin R;Chalmers MJ;Espada A;Dodge JA;Pascal BD;Griffin PR;Humblet C;Sali A
通讯作者：
Sali A
影响因子：
7.4
作者：
Sterling, Harry J.;Williams, Evan R.
通讯作者：
Williams, Evan R.
影响因子：
1.8
作者：
Liu J;McLuckey SA
通讯作者：
McLuckey SA
影响因子：
7.4
作者：
Zhang Z;Vachet RW
通讯作者：
Vachet RW