Trapped Ion Mobility Spectrometry of Native Macromolecular Assemblies,Analytical Chemistry

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Trapped Ion Mobility Spectrometry of Native Macromolecular Assemblies
Analytical Chemistry ( IF 6.7 ) Pub Date : 2021-01-25 , DOI: 10.1021/acs.analchem.0c04556
Kevin Jeanne Dit Fouque ₁ , Alyssa Garabedian ₁ , Fenfei Leng _{1,

2} , Yuk-Ching Tse-Dinh _{1,

2} , Mark E Ridgeway ₃ , Melvin A Park ₃ , Francisco Fernandez-Lima _{1,

2}

Affiliation

The structural elucidation of native macromolecular assemblies has been a subject of considerable interest in native mass spectrometry (MS), and more recently in tandem with ion mobility spectrometry (IMS-MS), for a better understanding of their biochemical and biophysical functions. In the present work, we describe a new generation trapped ion mobility spectrometer (TIMS), with extended mobility range (K₀ = 0.185–1.84 cm²·V^–1·s^–1), capable of trapping high-molecular-weight (MW) macromolecular assemblies. This compact 4 cm long TIMS analyzer utilizes a convex electrode, quadrupolar geometry with increased pseudopotential penetration in the radial dimension, extending the mobility trapping to high-MW species under native state (i.e., lower charge states). The TIMS capabilities to perform variable scan rate (Sr) mobility measurements over short time (100–500 ms), high-mobility resolution, and ion-neutral collision cross-section (CCS_N₂) measurements are presented. The trapping capabilities of the convex electrode TIMS geometry and ease of operation over a wide gas flow, rf range, and electric field trapping range are illustrated for the first time using a comprehensive list of standards varying from CsI clusters (n = 6–73), Tuning Mix oligomers (n = 1–5), common proteins (e.g., ubiquitin, cytochrome C, lysozyme, concanavalin (n = 1–4), carbonic anhydrase, β clamp (n = 1–4), topoisomerase IB, bovine serum albumin (n = 1–3), topoisomerase IA, alcohol dehydrogenase), IgG antibody (e.g., avastin), protein–DNA complexes, and macromolecular assemblies (e.g., GroEL and RNA polymerase (n = 1–2)) covering a wide mass (up to m/z 19 000) and CCS range (up to 22 000 Å² with <0.6% relative standard deviation (RSD)).

中文翻译：

天然大分子组装体的俘获离子淌度谱测定

天然大分子组装体的结构阐明一直是天然质谱 (MS) 领域备受关注的主题，最近又与离子迁移谱 (IMS-MS) 相结合，以更好地了解其生化和生物物理功能。在目前的工作中，我们描述了新一代俘获离子迁移谱仪（TIMS），其具有扩展的迁移率范围（ K ₀ = 0.185–1.84 cm ² ·V ^–1 ·s ^–1 ），能够捕获高分子量（ MW）大分子组装体。这款紧凑型 4 厘米长 TIMS 分析仪采用凸电极、四极几何形状，在径向维度上具有增强的赝电势穿透能力，将迁移率捕获扩展到自然状态（即较低电荷状态）下的高分子量物质。介绍了 TIMS 能够在短时间内（100–500 ms）执行可变扫描速率 ( Sr ) 迁移率测量、高迁移率分辨率和离子中性碰撞截面 (CCS _{N ₂} ) 测量。首次使用 CsI 簇的不同标准的综合列表（ n = 6–73）说明了凸电极 TIMS 几何形状的捕获能力以及在宽气流、射频范围和电场捕获范围内的易于操作性、Tuning Mix 寡聚体 ( n = 1–5)、常见蛋白质（例如泛素、细胞色素 C、溶菌酶、刀豆球蛋白 ( n = 1–4)、碳酸酐酶、β 钳 ( n = 1–4)、拓扑异构酶 IB、牛血清白蛋白（ n = 1–3）、拓扑异构酶 IA、乙醇脱氢酶）、IgG 抗体（例如阿瓦斯汀）、蛋白质-DNA 复合物和大分子组装体（例如、GroEL 和 RNA 聚合酶 ( n = 1–2)) 覆盖较宽的质量数（高达m / z 19 000）和 CCS 范围（高达 22 000 Å ² ，具有 <0.6% 相对标准偏差 (RSD)）。

更新日期：2021-02-09

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