Construction of a hybrid quadrupole/fourier transform ion cyclotron resonance mass spectrometer for versatile MS/MS above 10 kDa

Steven M. Patrie, Jay P. Charlebois, David Whipple, Neil L. Kelleher, Christopher L. Hendrickson, John P. Quinn, Alan G. Marshall, Biswarup Mukhopadhyay

Research output: Contribution to journalArticle

99 Scopus citations

Abstract

Technological advancements including an open-cylindrical Penning trap with capacitively coupled ICR cell, selective ion accumulation with a resolving quadrupole, and a voltage gradient used during ion extraction from an octopole ion trap, have individually improved dynamic range and sensitivity in Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR MS). Documented here is a new instrument utilizing these technologies toward the robust detection and fragmentation of biomolecules >10 kDa. Up to 55-fold enhancement in ion population by selective ion accumulation combined with 10- to 20- fold signal-to-noise improvement by application of a DC voltage gradient to an accumulation octopole during the ion transfer event offers improved signal-to-noise (or speed) of MS/MS experiments, for proteins from Methanococcus jannaschii and Saccharomyces cerevisiae whole cell lysates. After external quadrupole filtering with a 40 m/z window, three proteins were fragmented (and identified) in parallel from the database of Methanococcus jannaschii. Electron capture dissociation (ECD) of an intact yeast protein provides extensive sequence information resulting in a high degree of localization for an N-terminal acetylation. Hybrid fragmentation, infrared multiphoton dissociation (IRMPD) followed by low energy electrons (ECD), with the electron source located laterally off the z-axis and external to the magnet bore, presents a strategy for identification of proteins by means of the sequence tag approach. Automated implementation of diverse MS n approaches in a Q-FTMS instrument promises to help realize "top-down" proteomics in the future.

Original languageEnglish (US)
Pages (from-to)1099-1108
Number of pages10
JournalJournal of the American Society for Mass Spectrometry
Volume15
Issue number7
DOIs
StatePublished - Jul 1 2004

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy

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