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Ion Mobility Spectrometry-Hydrogen Deuterium Exchange Mass Spectrometry of Anions: Part 2. Assessing Charge Site Location and Isotope Scrambling

机译：离子的离子迁移谱-氢氘交换质谱法：第2部分。评估电荷位点位置和同位素加扰

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摘要

Ion mobility spectrometry (IMS) coupled with gas-phase hydrogen deuterium exchange (HDX)-mass spectrometry (MS) and molecular dynamic simulations (MDS) has been used for structural investigation of anions produced by electrospraying a sample containing a synthetic peptide having the sequence KKDDDDDIIKIIK. In these experiments the potential of the analytical method for locating charge sites on ions as well as for utilizing collision-induced dissociation (CID) to reveal the degree of deuterium uptake within specific amino acid residues has been assessed. For diffuse (i.e., more elongated) [M-2H]2− ions, decreased deuterium content along with MDS data suggest that the D4 and D6 residues are charge sites whereas for the more diffuse [M-3H]3− ions, the data suggest that the D4, D7, and the C-terminus are deprotonated. Fragmentation of mobility-selected, diffuse [M-2H]2− ions to determine deuterium uptake at individual amino acid residues reveals a degree of deuterium retention at incorporation sites. Although the diffuse [M-3H]3− ions may show more HD scrambling, it is not possible to clearly distinguish HD scrambling from the expected deuterium uptake based on a hydrogen accessibility model. The capability of the IMS-HDX-MS/MS approach to provide relevant details about ion structure is discussed. Additionally, the ability to extend the approach for locating protonation sites on positively-charged ions is presented.
机译：离子淌度光谱（IMS）结合气相氢氘交换（HDX）-质谱（MS）和分子动力学模拟（MDS）已用于对通过电喷雾包含具有以下序列的合成肽的样品产生的阴离子进行结构研究KKDDDDDIIKIIK。在这些实验中，已评估了分析方法在离子上定位电荷位点以及利用碰撞诱导解离（CID）揭示特定氨基酸残基中氘吸收程度的潜力。对于弥散的（即更伸长的）[M-2H] 2-离子，氘含量的降低以及MDS数据表明D4和D6残基是电荷位点，而对于弥散的[M-3H] ] 3-离子，数据表明D4，D7和C端均已去质子化。通过迁移率选择的，分散的[M-2H] 2-离子的碎裂来确定各个氨基酸残基的氘吸收，揭示了氘在结合位点的保留程度。尽管扩散的[M-3H] 3-离子可能显示更多的HD加扰，但无法基于氢可及性模型将HD加扰与预期的氘吸收区分开来。讨论了IMS-HDX-MS / MS方法提供有关离子结构的相关详细信息的能力。此外，还提供了扩展在正电荷离子上定位质子化位点的方法的能力。

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