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Activated Ion Electron Capture Dissociation (AI ECD) of proteins: synchronization of infrared and electron irradiation with ion magnetron motion.

Mikhailov, Victor A. and Cooper, Helen J. (2008) Activated Ion Electron Capture Dissociation (AI ECD) of proteins: synchronization of infrared and electron irradiation with ion magnetron motion. Journal of the American Society for Mass Spectrometry, 20 (5). pp. 763-771. ISSN 1044-0305

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URL of Published Version: http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=19200749

Identification Number/DOI: doi: 10.1016/j.jasms.2008.12.015

Here, we show that to perform activated ion electron capture dissociation (AI-ECD) in a Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer equipped with a CO(2) laser, it is necessary to synchronize both infrared irradiation and electron capture dissociation with ion magnetron motion. This requirement is essential for instruments in which the infrared laser is angled off-axis, such as the Thermo Finnigan LTQ FT. Generally, the electron irradiation time required for proteins is much shorter (ms) than that required for peptides (tens of ms), and the modulation of ECD, AI ECD, and infrared multiphoton dissociation (IRMPD) with ion magnetron motion is more pronounced. We have optimized AI ECD for ubiquitin, cytochrome c, and myoglobin; however the results can be extended to other proteins. We demonstrate that pre-ECD and post-ECD activation are physically different and display different kinetics. We also demonstrate how, by use of appropriate AI ECD time sequences and normalization, the kinetics of protein gas-phase refolding can be deconvoluted from the diffusion of the ion cloud and measured on the time scale longer than the period of ion magnetron motion.

Type of Work:Article
Date:31 December 2008 (Publication)
School/Faculty:Schools (1998 to 2008) > School of Biosciences
Department:Structural Biology
Subjects:Q Science (General)
Institution:University of Birmingham
Copyright Holders:Elsevier
ID Code:240
Refereed:YES
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