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Title: Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis

Abstract

High resolution mass spectrometry is a key technology for in-depth protein characterization. High-field Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) enables high-level interrogation of intact proteins in the most detail to date. However, an appropriate complement of fragmentation technologies must be paired with FTMS to provide comprehensive sequence coverage, as well as characterization of sequence variants, and post-translational modifications. Here we describe the integration of front-end electron transfer dissociation (FETD) with a custom-built 21 tesla FT-ICR mass spectrometer, which yields unprecedented sequence coverage for proteins ranging from 2.8 to 29 kDa, without the need for extensive spectral averaging (e.g., ~60% sequence coverage for apo-myoglobin with four averaged acquisitions). The system is equipped with a multipole storage device separate from the ETD reaction device, which allows accumulation of multiple ETD fragment ion fills. Consequently, an optimally large product ion population is accumulated prior to transfer to the ICR cell for mass analysis, which improves mass spectral signal-to-noise ratio, dynamic range, and scan rate. We find a linear relationship between protein molecular weight and minimum number of ETD reaction fills to achieve optimum sequence coverage, thereby enabling more efficient use of instrument data acquisition time. Finally, real-time scaling of the numbermore » of ETD reactions fills during method-based acquisition is shown, and the implications for LC-MS/MS top-down analysis are discussed. .« less

Authors:
;  [1]; ; ; ;  [2];  [3]; ;  [1];  [3]; ;  [1];  [3]
  1. National High Magnetic Field Laboratory (NHMFL) (United States)
  2. Thermo Fisher Scientific (United States)
  3. University of Virginia, Department of Chemistry (United States)
Publication Date:
OSTI Identifier:
22776860
Resource Type:
Journal Article
Journal Name:
Journal of the American Society for Mass Spectrometry
Additional Journal Information:
Journal Volume: 28; Journal Issue: 9; Other Information: Copyright (c) 2017 American Society for Mass Spectrometry; http://www.springer-ny.com; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 1044-0305
Country of Publication:
United States
Language:
English
Subject:
46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND TECHNOLOGY; DATA ACQUISITION; DISSOCIATION; ELECTRON TRANSFER; EQUIPMENT; FOURIER TRANSFORMATION; FRAGMENTATION; ION CYCLOTRON-RESONANCE; IONIZATION; MASS SPECTROMETERS; MASS SPECTROSCOPY; MOLECULAR WEIGHT; SIGNAL-TO-NOISE RATIO

Citation Formats

Weisbrod, Chad R., E-mail: weisbrod@magnet.fsu.edu, Kaiser, Nathan K., Syka, John E. P., Early, Lee, Mullen, Christopher, Dunyach, Jean-Jacques, English, A. Michelle, Anderson, Lissa C., Blakney, Greg T., Shabanowitz, Jeffrey, Hendrickson, Christopher L., Marshall, Alan G., and Hunt, Donald F. Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis. United States: N. p., 2017. Web. doi:10.1007/S13361-017-1702-3.
Weisbrod, Chad R., E-mail: weisbrod@magnet.fsu.edu, Kaiser, Nathan K., Syka, John E. P., Early, Lee, Mullen, Christopher, Dunyach, Jean-Jacques, English, A. Michelle, Anderson, Lissa C., Blakney, Greg T., Shabanowitz, Jeffrey, Hendrickson, Christopher L., Marshall, Alan G., & Hunt, Donald F. Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis. United States. doi:10.1007/S13361-017-1702-3.
Weisbrod, Chad R., E-mail: weisbrod@magnet.fsu.edu, Kaiser, Nathan K., Syka, John E. P., Early, Lee, Mullen, Christopher, Dunyach, Jean-Jacques, English, A. Michelle, Anderson, Lissa C., Blakney, Greg T., Shabanowitz, Jeffrey, Hendrickson, Christopher L., Marshall, Alan G., and Hunt, Donald F. Fri . "Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis". United States. doi:10.1007/S13361-017-1702-3.
@article{osti_22776860,
title = {Front-End Electron Transfer Dissociation Coupled to a 21 Tesla FT-ICR Mass Spectrometer for Intact Protein Sequence Analysis},
author = {Weisbrod, Chad R., E-mail: weisbrod@magnet.fsu.edu and Kaiser, Nathan K. and Syka, John E. P. and Early, Lee and Mullen, Christopher and Dunyach, Jean-Jacques and English, A. Michelle and Anderson, Lissa C. and Blakney, Greg T. and Shabanowitz, Jeffrey and Hendrickson, Christopher L. and Marshall, Alan G. and Hunt, Donald F.},
abstractNote = {High resolution mass spectrometry is a key technology for in-depth protein characterization. High-field Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) enables high-level interrogation of intact proteins in the most detail to date. However, an appropriate complement of fragmentation technologies must be paired with FTMS to provide comprehensive sequence coverage, as well as characterization of sequence variants, and post-translational modifications. Here we describe the integration of front-end electron transfer dissociation (FETD) with a custom-built 21 tesla FT-ICR mass spectrometer, which yields unprecedented sequence coverage for proteins ranging from 2.8 to 29 kDa, without the need for extensive spectral averaging (e.g., ~60% sequence coverage for apo-myoglobin with four averaged acquisitions). The system is equipped with a multipole storage device separate from the ETD reaction device, which allows accumulation of multiple ETD fragment ion fills. Consequently, an optimally large product ion population is accumulated prior to transfer to the ICR cell for mass analysis, which improves mass spectral signal-to-noise ratio, dynamic range, and scan rate. We find a linear relationship between protein molecular weight and minimum number of ETD reaction fills to achieve optimum sequence coverage, thereby enabling more efficient use of instrument data acquisition time. Finally, real-time scaling of the number of ETD reactions fills during method-based acquisition is shown, and the implications for LC-MS/MS top-down analysis are discussed. .},
doi = {10.1007/S13361-017-1702-3},
journal = {Journal of the American Society for Mass Spectrometry},
issn = {1044-0305},
number = 9,
volume = 28,
place = {United States},
year = {2017},
month = {9}
}