Abstract
Thirty years after its invention, the Fourier transform ion cyclotron resonance (FTICR) mass spectrometer continues its evolution into an enabling technology to deepen our understanding of biological systems. For contemporary protein analysis, a quadrupole FTICR hybrid with electrospray ionization is forwarded here as an engine for high resolution tandem mass spectrometry in a high-throughput setting. Three basic strategies for MS/MS of ions >10 kDa are illustrated by identification and characterization of proteins from stationary-phase yeast cells. From samples containing 1-13 proteins, introduced by a nanospray robot, 2-6 can be isolated automatically and fragmented in 15-45 min. Features used commonly for peptide analyses (e.g., multidimensional separations, data dependent acquisition, and probability-based protein identification) are now available in an off-line top-down platform. In one set of 9 samples, 20 proteins (6-17 kDa) were processed through the platform yielding a mean Pscore=0.002 (99.8% identification confidence) upon database retrieval with characterization of N-terminal post-translational modifications. On occasion of his 60th birthday, we offer this work in celebration of the steadily advancing technology that Alan Marshall helped to invent (now in over 400+ labs worldwide).
Original language | English (US) |
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Pages (from-to) | 175-184 |
Number of pages | 10 |
Journal | International Journal of Mass Spectrometry |
Volume | 234 |
Issue number | 1-3 |
DOIs | |
State | Published - May 1 2004 |
Funding
We thank many members of the Kelleher Group for technical assistance and helpful discussions. The authors acknowledge the National Science Foundation (CHE-0134953), the National Institutes of Health (GM 067193), and the Packard and Sloan Foundations. Finally, we thank Alan Marshall, Chris Hendrickson, John Quinn, and Greg Blakney for their gracious support of our FTICR MS construction project, made possible by the NSF National High-Field FTICR Mass Spectrometry Facility (CHE 99-09502), Florida State University, and the National High Magnetic Field Laboratory in Tallahassee, Florida.
Keywords
- FTICR
- FTMS
- Proteomics
- Top-down
ASJC Scopus subject areas
- Instrumentation
- Condensed Matter Physics
- Spectroscopy
- Physical and Theoretical Chemistry