Improving Ion Mobility Mass Spectrometry of Proteins through Tristate Gating and Optimization of Multiplexing Parameters
Improving Ion Mobility Mass Spectrometry of Proteins through Tristate Gating and Optimization of Multiplexing Parameters
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DOI：
10.1021/jasms.2c00274
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发表时间：
2022-12-05
影响因子：
3.2
通讯作者：
Brodbelt, Jennifer S.
中科院分区：
文献类型：
--
作者：
Butalewicz, Jamie P.;Sanders, James D.;Brodbelt, Jennifer S.
将漂移管离子迁移率（IM）与傅里叶变换质谱（FT-MS）耦合，为基于大小和构象的离子气相分离提供了高分辨率质量分析的机会。然而，结合IM和FT-MS是具有挑战性的，因为离子离开漂移管的时间尺度比质量分析的速度快得多。傅立叶变换（FT）和哈达玛变换复用方法已经实现，以克服占空比不匹配，为获得高分辨率，高质量精度的迁移率选择离子的分析提供了新的途径。由于众所周知的栅极损耗效应，用于集成漂移管和FT质量分析仪的门控方法对大的、低迁移率离子的传输进行了区分。三态门控策略已被证明可以增加漂移管IM-FT-MS系统的离子传输，通过实施双离子门控，控制离子通过漂移管的数量和时间，以减少缓慢移动离子的损失。在这里，我们提出了一套优化的多路复用参数，用于几种蛋白质在Orbitrap质谱仪上的三态门控离子迁移，并进一步报告了增加离子传输和迁移分辨率以及将实验时间从15分钟减少到30秒的参数。在到达时间分布（ATDs）中，泛素的峰值强度平均增加2.1倍，肌红蛋白的峰值强度平均增加1.5倍，分辨力平均增加11%。
Coupling drift tube ion mobility (IM) to Fourier transform mass spectrometry (FT-MS) affords the opportunity for gas-phase separation of ions based on size and conformation with high-resolution mass analysis. However, combining IM and FT-MS is challenging because ions exit the drift tube on a much faster time scale than the rate of mass analysis. Fourier transform (FT) and Hadamard transform multiplexing methods have been implemented to overcome the duty-cycle mismatch, offering new avenues for obtaining high-resolution, high-mass-accuracy analysis of mobility-selected ions. The gating methods used to integrate the drift tube with the FT mass analyzer discriminate against the transmission of large, low-mobility ions owing to the well-known gate depletion effect. Tristate gating strategies have been shown to increase ion transmission for drift tube IM-FT-MS systems through implementation of dual ion gating, controlling the quantity and timing of ions through the drift tube to reduce losses of slow-moving ions. Here we present an optimized set of multiplexing parameters for tristate gating ion mobility of several proteins on an Orbitrap mass spectrometer and further report parameters for increased ion transmission and mobility resolution as well as decreased experimental times from 15 min down to 30 s. On average, peak intensities in the arrival time distributions (ATDs) for ubiquitin increased 2.1x on average, while those of myoglobin increased by 1.5x with a resolving power increase on average of 11%.