Improving the Performance of a Quadrupole Time-of-Flight Instrument for Macromolecular Mass Spectrometry

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• 作者：Robert H. H. van den Heuvel ; Esther van Duijn ; Hortense Mazon ; Silvia A. Synowsky ; Kristina Lorenzen ; Cees Versluis ; Stan J. J. Brouns ; Dave Langridge ; John van der Oost ; John Hoyes ; Albert J. R. Heck
• 刊名：Analytical Chemistry
• 出版年：2006
• 出版时间：November 1, 2006
• 年：2006
• 卷：78
• 期：21
• 页码：7473 - 7483
• 全文大小：459K
• 年卷期：v.78,no.21(November 1, 2006)
• ISSN：1520-6882

文摘

We modified and optimized a first generation quadrupoletime-of-flight (Q-TOF) 1 to perform tandem mass spectrometry on macromolecular protein complexes. Themodified instrument allows isolation and subsequentdissociation of high-mass protein complexes throughcollisions with argon molecules. The modifications of theQ-TOF 1 include the introduction of (1) a flow-restrictingsleeve around the first hexapole ion bridge, (2) a low-frequency ion-selecting quadrupole, (3) a high-pressurehexapole collision cell, (4) high-transmission grids in themulticomponent ion lenses, and (5) a low repetition ratepusher. Using these modifications, we demonstrate theexperimental isolation of ions up to 12 800 mass-to-charge units and detection of product ions up to 38 150Da, enabling the investigation of the gas-phase stability,protein complex topology, and quaternary structure ofprotein complexes. Some of the data reveal a so-farunprecedented new mechanism in gas-phase dissociationof protein oligomers whereby a tetramer complex dissociates into two dimers. These data add to the current debatewhether gas-phase structures of protein complexes doretain some of the structural features of the correspondingspecies in solution. The presented low-cost modificationson a Q-TOF 1 instrument are of interest to everyoneworking in the fields of macromolecular mass spectrometry and more generic structural biology.