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ECD of Tyrosine Phosphorylation in a Triple Quadrupole Mass Spectrometer with a Radio-Frequency-Free Electromagnetostatic Cell

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Journal of The American Society for Mass Spectrometry

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Abstract

A radio frequency-free electromagnetostatic (EMS) cell devised for electron-capture dissociation (ECD) of ions has been retrofitted into the collision-induced dissociation (CID) section of a triple quadrupole mass spectrometer to enable recording of ECD product-ion mass spectra and simultaneous recording of ECD-CID product-ion mass spectra. This modified instrument can be used to produce easily interpretable ECD and ECD-CID product-ion mass spectra of tyrosine-phosphorylated peptides that cover over 50% of their respective amino-acid sequences and readily identify their respective sites of phosphorylation. ECD fragmentation of doubly protonated, tyrosine-phosphorylated peptides, which was difficult to observe with FT-ICR instruments, occurs efficiently in the EMS cell.

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Acknowledgments

Various aspects of this research were supported by grants from the NSF (CHE-0924027), the Oregon Nanoscience and Microtechnologies Institute (#09-31 #3.5), NIH NCRR (R01RR026275), NIH NIEHS (ES00210–Environmental Health Sciences Center), and Agilent Technologies (#2443). The authors thank Peter D. Hoffman (Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, Oregon) for his assistance in collecting some of the data presented in this manuscript, Elsworth T. Hinke and Larry M. Nelson (Department of Chemistry, Oregon State University, Corvallis, Oregon) for their assistance in fabricating components of the EMS ECD cell used in this study, Piriya Wongkongkathep and Dr. Joseph Loo (Department of Chemistry and Biochemistry, University of California-Los Angeles) for graciously acquiring and labeling the Bruker 15 T SolariX ECD FT-ICR product-ion mass spectra displayed in this report, and George Stafford (Agilent Technologies, Santa Clara, California) for generously reading the manuscript and offering several useful suggestions for improving its content.

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Authors and Affiliations

  1. Department of Chemistry, Oregon State University, Corvallis, OR, 97333, USA

    Valery G. Voinov & Douglas F. Barofsky

  2. Department of Environmental and Molecular Toxicology, Oregon State University, Corvallis, OR, 97333, USA

    Samuel E. Bennett

  3. Department of Biochemistry and Biophysics, Oregon State University, Corvallis, OR, 97333, USA

    Joseph S. Beckman

  4. Pacific Institute of Bioorganic Chemistry, Vladivostok, Russia

    Valery G. Voinov

Authors
  1. Valery G. Voinov
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  2. Samuel E. Bennett
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  3. Joseph S. Beckman
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  4. Douglas F. Barofsky
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Corresponding author

Correspondence to Valery G. Voinov.

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Voinov, V.G., Bennett, S.E., Beckman, J.S. et al. ECD of Tyrosine Phosphorylation in a Triple Quadrupole Mass Spectrometer with a Radio-Frequency-Free Electromagnetostatic Cell. J. Am. Soc. Mass Spectrom. 25, 1730–1738 (2014). https://doi.org/10.1007/s13361-014-0956-2

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  • DOI: https://doi.org/10.1007/s13361-014-0956-2

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