Improving the performance of a quadrupole time-of-flight instrument for macromolecular mass spectrometry
Improving the performance of a quadrupole time-of-flight instrument for macromolecular mass spectrometry
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DOI：
10.1021/ac061039a
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发表时间：
2006-11-01
影响因子：
7.4
通讯作者：
Heck, Albert J. R.
中科院分区：
文献类型：
--
作者：
van den Heuvel, Robert H. H.;van Duijn, Esther;Heck, Albert J. R.
我们修改和优化了第一代四极杆飞行时间（Q-TOF）1进行串联质谱的大分子蛋白质复合物。改进的仪器允许通过与氩分子的碰撞分离和随后的高质量蛋白质复合物的解离。Q-TOF 1的修改包括引入（1）围绕第一六极离子桥的限流套管，（2）低频离子选择四极，（3）高压六极碰撞室，（4）多组分离子透镜中的高透射栅格，以及（5）低重复率推动器。使用这些修改，我们证明了实验隔离的离子高达12 800质量-电荷单位和检测的产品离子高达38 150 Da，使调查的气相稳定性，蛋白质复合物的拓扑结构，和四级结构的蛋白质复合物。一些数据揭示了一个迄今为止前所未有的新机制，在气相解离的蛋白质低聚物，其中一个四聚体复合物解离成两个二聚体。这些数据增加了目前的争论是否气相结构的蛋白质复合物保留了一些结构特征的相应物种在溶液中。Q-TOF 1仪器上的低成本修改对大分子质谱和更通用的结构生物学领域的每个人都很感兴趣。
We modified and optimized a first generation quadrupole time-of-flight (Q-TOF) 1 to perform tandem mass spectrometry on macromolecular protein complexes. The modified instrument allows isolation and subsequent dissociation of high-mass protein complexes through collisions with argon molecules. The modifications of the Q-TOF 1 include the introduction of (1) a flow-restricting sleeve around the first hexapole ion bridge, (2) a low-frequency ion-selecting quadrupole, (3) a high-pressure hexapole collision cell, (4) high-transmission grids in the multicomponent ion lenses, and (5) a low repetition rate pusher. Using these modifications, we demonstrate the experimental isolation of ions up to 12 800 mass-to-charge units and detection of product ions up to 38 150 Da, enabling the investigation of the gas-phase stability, protein complex topology, and quaternary structure of protein complexes. Some of the data reveal a so-far unprecedented new mechanism in gas-phase dissociation of protein oligomers whereby a tetramer complex dissociates into two dimers. These data add to the current debate whether gas-phase structures of protein complexes do retain some of the structural features of the corresponding species in solution. The presented low-cost modifications on a Q-TOF 1 instrument are of interest to everyone working in the fields of macromolecular mass spectrometry and more generic structural biology.