Journal of the American Society for Mass Spectrometry

, Volume 21, Issue 12, pp 1966–1968

Native electrospray and electron-capture dissociation in FTICR mass spectrometry provide top-down sequencing of a protein component in an intact protein assembly

Authors

  • Hao Zhang
    • Department of ChemistryWashington University in St. Louis
  • Weidong Cui
    • Department of ChemistryWashington University in St. Louis
  • Jianzhong Wen
    • Department of ChemistryWashington University in St. Louis
  • Robert E. Blankenship
    • Department of ChemistryWashington University in St. Louis
    • Department of ChemistryWashington University in St. Louis
Short Comunication

DOI: 10.1016/j.jasms.2010.08.006

Cite this article as:
Zhang, H., Cui, W., Wen, J. et al. J Am Soc Mass Spectrom (2010) 21: 1966. doi:10.1016/j.jasms.2010.08.006

Abstract

The intact yeast alcohol dehydrogenase (ADH) tetramer of 147 kDa was introduced into a FTICR mass spectrometer by native electrospray. Electron capture dissociation of the entire 23+ to 27+ charge state distribution produced the expected charge-reduced ions and, more unexpectedly, 39 c-type peptide fragments that identified N-terminus acetylation and the first 55 amino acids. The results are in accord with the crystal structure of yeast ADH, which shows that the C-terminus is buried at the assembly interface, whereas the N-terminus is exposed, allowing ECD to occur. This remarkable observation shows promise that a top-down approach for intact protein assemblies will be effective for characterizing their components, inferring their interfaces, and obtaining both proteomics and structural biology information in one experiment.

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© American Society for Mass Spectrometry. Published by Elsevier Inc 2010