Improving the performance of a quadrupole time-of-flight instrument for macromolecular mass spectrometry
| Autor: | Kristina Lorenzen, John van der Oost, Hortense Mazon, Dave Langridge, Stan J. J. Brouns, Esther van Duijn, A.J.R. Heck, Robert H. H. van den Heuvel, John Hoyes, Cees Versluis, Silvia A. Synowsky |
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| Přispěvatelé: | Biomoleculaire Massaspectrometrie, Massaspectrometrie, Dep Scheikunde |
| Rok vydání: | 2006 |
| Předmět: |
infrared radiative dissociation
Electrospray ionization Analytical chemistry Mass spectrometry Tandem mass spectrometry vanillyl-alcohol oxidase 01 natural sciences Microbiology Dissociation (chemistry) Algemeen onderzoek Analytical Chemistry Ion Farmacie/Biofarmaceutische wetenschappen (FARM) 03 medical and health sciences Microbiologie Tandem Mass Spectrometry Pathology Chaperonin 10 Molecule Geneeskunde(GENK) 030304 developmental biology VLAG 0303 health sciences protein complexes Chemistry Geneeskunde (GENK) 010401 analytical chemistry Farmacie(FARM) Chaperonin 60 assemblies electrospray-ionization General [Econometric and Statistical Methods] groel 0104 chemical sciences Alcohol Oxidoreductases escherichia-coli strain esi-ms Quadrupole ions Protein quaternary structure gas-phase dissociation |
| Zdroj: | Analytical Chemistry 78 (2006) 21 Analytical Chemistry, 78(21), 7473-7483 |
| ISSN: | 0003-2700 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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