ETD in a traveling wave ion guide at tuned Z-spray ion source conditions allows for site-specific hydrogen/deuterium exchange measurements.
ETD in a traveling wave ion guide at tuned Z-spray ion source conditions allows for site-specific hydrogen/deuterium exchange measurements.
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DOI：
10.1007/s13361-011-0196-7
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发表时间：
2011-10
影响因子：
3.2
通讯作者：
Brown, Jeffery M.
中科院分区：
文献类型：
--
作者：
Rand, Kasper D.;Pringle, Steven D.;Morris, Michael;Engen, John R.;Brown, Jeffery M.
关键词：
最近应用电子转移解离（ETD）在单残基分辨率（HX-ETD）下测量溶液中蛋白质的氢交换，为基于质谱的生物分子结构分析铺平了道路，达到了前所未有的详细水平。该方法要求在ETD之前多肽离子的激活最小，以防止溶液中氘标签的不良气相随机化（即氢乱序）。在这里，我们探索了在四极飞行时间（Q-TOF）质谱仪的行波离子波导中使用“z喷雾”型离子源的ETD来测量肽中单个残基的氘含量。我们系统地确定了有助于碰撞激活的z喷射离子源的关键参数，并确定了允许在行波离子波导中进行无气相氢扰的ETD实验的条件。我们发现，在随后的行波离子波导中，ETD和补充碰撞激活可以改善低电荷肽中残基特异性氘含量的提取。我们的结果证明了这种方法的可行性，并说明了在配备行波离子波导的高分辨率Q-TOF仪器上进行HX-ETD实验的优势。有助于离子加热的z喷射离子源参数的确定同样与越来越多的质谱应用相关，这些应用也依赖于离子向气相的能量温和转移，例如通过结合气相离子-离子/离子中性反应或离子迁移谱法的天然质谱法分析生物分子结构。
The recent application of electron transfer dissociation (ETD) to measure the hydrogen exchange of proteins in solution at single-residue resolution (HX-ETD) paves the way for mass spectrometry-based analyses of biomolecular structure at an unprecedented level of detail. The approach requires that activation of polypeptide ions prior to ETD is minimal so as to prevent undesirable gas-phase randomization of the deuterium label from solution (i.e., hydrogen scrambling). Here we explore the use of ETD in a traveling wave ion guide of a quadrupole-time-of-flight (Q-TOF) mass spectrometer with a “Z-spray” type ion source, to measure the deuterium content of individual residues in peptides. We systematically identify key parameters of the Z-spray ion source that contribute to collisional activation and define conditions that allow ETD experiments to be performed in the traveling wave ion guide without gas-phase hydrogen scrambling. We show that ETD and supplemental collisional activation in a subsequent traveling wave ion guide allows for improved extraction of residue-specific deuterium contents in peptides with low charge. Our results demonstrate the feasibility, and illustrate the advantages of performing HX-ETD experiments on a high-resolution Q-TOF instrument equipped with traveling wave ion guides. Determination of parameters of the Z-spray ion source that contribute to ion heating are similarly pertinent to a growing number of MS applications that also rely on an energetically gentle transfer of ions into the gas-phase, such as the analysis of biomolecular structure by native mass spectrometry in combination with gas-phase ion-ion/ion-neutral reactions or ion mobility spectrometry.
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