Analysis of Native-Like Ions Using Structures for Lossless Ion Manipulations
Analysis of Native-Like Ions Using Structures for Lossless Ion Manipulations
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DOI：
10.1021/acs.analchem.6b02089
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发表时间：
2016-09-20
影响因子：
7.4
通讯作者：
Bush, Matthew F.
中科院分区：
文献类型：
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作者：
Allen, Samuel J.;Eaton, Rachel M.;Bush, Matthew F.
天然类蛋白质和蛋白质络合物离子的离子迁移率分离扩展了天然质谱分析的结构信息。在这里，我们实现了用于分析类自然离子的无损离子操纵结构(SLIM)。此前，SILM已经被证明在质量高达3000 Da的离子几乎无损传输的情况下工作。首次使用SLIM分离了质量从12 kDa到145 kDa的类天然蛋白质和蛋白质络合物离子。由此得到的到达时间分布是单峰的，并被用来确定碰撞截面值，这些值在射频限制漂移室测量所确定的值的3%以内。这些结果与这些实验中保留的类自然离子结构是一致的。用SLIM测得的类天然离子的表观分辨能力高达42，这是直接从实验数据中报道的蛋白质络合物类天然离子的最高值。有趣的是，表观分辨能力强烈地依赖于分析物的同一性，这表明这些离子的到达时间分布可能与每个分析物独特的气相结构集合有关。这些结果表明。新出现的超薄技术的广泛范围可能都适用于类天然离子的分析，这将使未来在结构生物学、生物物理学和生物制药表征领域的应用成为可能。
Ion mobility separation of native-like protein and protein complex ions expands the structural information available through native mass spectrometry analysis. Here, we implement Structures for Lossless Ion Manipulations (SLIM) for the analysis of native-like ions. SLIM has been shown previously to operate with near lossless transmission of ions up to 3000 Da in mass. Here for the first time, SLIM was used to separate native-like protein and protein complex ions ranging in mass from 12 to 145 kDa. The resulting arrival-time distributions were monomodal and were used to determine collision cross section values that are within 3% of those determined from radio-frequency-confining drift cell measurements. These results are consistent with the retention of native-like ion structures throughout these experiments. The apparent resolving powers of native-like ions measured using SLIM are as high as 42, which is the highest value reported directly from experimental data for the native-like ion of a protein complex. Interestingly, the apparent resolving power depends strongly on the identity of the analyte, suggesting that the arrival-time distributions of these ions may have contributions from an ensemble of structures in the gas phase that is unique to each analyte. These results suggest. hat the broad range of emerging SLIM technologies may all be adaptable to the analysis of native-like ions, which will enable future applications in the areas of structural biology; biophysics, and biopharmaceutical characterization.