期刊
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
卷 276, 期 2-3, 页码 136-143出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.ijms.2008.07.029
关键词
ion-ion reaction; ion parking; derivatized protein; high charge state; ion attachment
资金
- NIH Biotechnology Training
- National Heart Lung Blood Institute (NHLBI) [N01-HV-28182]
- NIH [1R01GM080148, R01GM065406, R33DK070297]
- NSF [0701846, 0747990]
- University of Wisconsin-Madison, Thermo Fisher
- Beckman Foundation
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [0747990] Funding Source: National Science Foundation
Electrospray ionization (ESI) of denatured proteins produces a mass spectrum with a broad distribution of multiply charged ions. Attaching fixed positive charges, specifically quaternary ammonium groups, to proteins at their carboxylic acid groups generates substantially higher charge states compared to the corresponding unmodified proteins in positive-mode ESI. Ion-ion reactions of these modified proteins with reagent anions leads to charge reduction by proton transfer. These proton transfer reactions cannot remove charge from the quaternary ammonium groups, which do not have a proton to transfer to the anion. Thus, one might expect charge reduction to stop at a single charge state equal to the number of fixed charges on the modified protein. However, ion-ion reactions yield charge states lower than this number of fixed charges due to anion attachment (adduction) to the proteins. Charge reduction via ion-molecule reactions involving gas-phase bases also give adducts on the modified protein ions in low charge states. Such adducts are avoided by keeping the ions in charge states well above the number of fixed charges. In the present work protein ions were selectively parked within an ion trap mass spectrometer in a high charge state by mild radiofrequency excitation that dramatically slows their ion-ion reaction rate-a technique termed ion parking. The combination of ion parking with the fixed-charge modified proteins permits generation of a large population of ions in a single, very high charge state. (C) 2008 Elsevier B.V. All rights reserved.
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